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JP WO2007/108559 A1 2007.9.27 10 (57)【要約

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5. 000000000000000000000 000 Hod 100000000000000000000000 100000000000000000000000 000
6. 02007 108559 Al 2007 9 27 132 c D
7. 200000000000000000000000000000000000 5929 7010 5 10 2 10 1 6 Su 990000000 1000000000000000003000000000000 100000000000000000 10000 0 FMA
8. 6 1963344 IlgG3344h1 0000 1gG3344 N5KG3331 000000 PCRI 00000060 Overlap Extention HH 100 N5KG3331 GGG4 1 P2 00 O 940 10 94C 15 55 100
9. N5KG4PEH HHHHHHHHHHHHHHHHHHHHHHHHHH gG4PE 00000 00000000 CMI 000 19891 70 5 UK2211504 0 40 SV40 HHHHHHHHHHHHHHHHHHH M m EM RT E 40 bp100 2709 000
10. 1000000 10000000000 10 1960000000 00000000000 1 19 0 0 4 49 19610 0 7 10 1gG1H 7 10 4 49 00 7 10 H 4 499 0000 0000000000000000000 5 Sallg 3 Nhel 1000000000000000000000 000 0 0 0 0 KOD Plus DN D D PCR 0 00000 1000000000000000000000
11. O 00000000000000000000 100000000000000000000000000 0 000 7 1000 000 00 7 10G4 4uhm 01 0 O 000110 class HLA DR DQ DP TH TCR 100000 THHHHHHHHHHHHHHHHHHHHHHHHH
12. _R20 5 AGAGAGAGAG CGGCCGCTCA AGGCTGCTGC GTG 3 00000 130 ML3 20FW 5 GTAAAACGACGGCCAGTG 3 ML3RV 5 CAGGAAACAGCTATGAC 3 0 0 0 O 150 2 000000 0 6 TPO hMIH 000 5080 Trp Ser 0 Abe Leukem 2002 Aug 16 8 1500 1506 DO l li 5080 Toe 0000 pEF MPL6350 GeneEditor in vitro Site Directed Mutagenesi s System UUULHULELBHULBUtHIEEglidglidutlitluiiultiil 5
13. 0 0 UI TPO 1000 100 2 1003 100500 00 7 1000 8 100 1000 1000 100000000000000 1000000000000000 100 HHHHHHHHHHHHH Genban 1000 000000000 000000 000000 100000000000000 00000000000 100000000000000 100000000000000 20 0000 BS 3 228 341 10000010 00000000 1000000000000000 100000000000000 cm 0000000000000 136 WO2007 108559 1 2007
14. 10 Henatracer801 ADPH HHHHHHHHHHHHIH ADPG 00000 10000000000000000000000000000 Po 100000000 PEG rHuMGDFH 10 g kgH 10000000000000000000000 7 106PEG 0 7 106334411 0 0 0 0 0 199 00000000 A BI J O 7 1063344111 00000 HHHHHHHHH 10 100000000000000000000000000 HHHHHHHHHHHHH nn NOG NOD SCID IL2 y
15. 10000000000000000000000000000 1000000000000000000000000000 YHHHHHHHHHHHHHHHHHHHHHHHH 100000000000000000 1965100000000 N5KG4PE DEC Pharmaceuticals 19G4PEH 19G4PEH 19G4H Ser228Pro Leu235Gl ul Ser228Pro Leu235G ul 0 Antibody dependent cellular cytotoxicity ADCO O N5KG1 19610 0 0 0 0 19630 N5KG3H N5KG1 N5KG3 HHHHHHHHHHHHHH
16. 1000000000000 10 20 30 40 50 9 02007 108559 1 2007 9 27
17. 47 02007 108559 Al 2007 9 27 100000000000000000 100000000000000000 0000000 0 0000000000000000000000000000 a za ERANO RS RES T EM E C NEM C C C C C Em C mm eC ri non C x5 C C C C C C r1 C r1 r1 C C r C 17 18 5 AGGTGCTGGG CACCGTGGGC ATGTGTGAGT 3 70 17 2F 5 CACACATGCC CACGGTGCCC AGCACCTGAG TTC 3 71 nn oo C r1 C C r1 G C i E C Xu EDS E a 3 H n n n C r1 an C r1 10 20 30
18. 10000000000000000000000000000 1000000 0 10 1 ITPH 2 000 3 4 5 O 6 1000000000000 10000000000000000000000000000 100000000000000000000000000000000 00 200 100 270 2008 10 27 n 1 0 3 1 2 100000000 3 100 2 100000000 100000000 ELKTPLGDTTHT 10 0 0 CPRCP EPKSCDTPPPCP RCP x30 CPRCPEPKSCDTPPPCPRCP CPRCPH ESKTPLGDTTHI 000 11 000 000000 100000000000 000 b CD34 CFU 0000 0 10 000ng
19. J 00344 000000 00 1 000ng 5 UT7 H PEGrHuMGDFH 70 EC50 10nM O dE E ER DICERE OD O00 0 0 b a CD34H 000000 0000000000000 10019 mg 00 100000000 b UT7 TPO PEGrHuMGDF 90 0000 EC50 1nM 100000000 1 8 1 2 3 4 uu 5 6 7 8 000 1000 1100 2 1 1
20. 10 20 30 40 50 22 P WO2007 108559 Al 2007 9 27 eu h ATE Eu cut NEE 00000000 DIRE 0000000000000000000000000000000 000 2 HHHHHHHHHHHHHHHH HHHHHHHHHHHHHHH LX RR HHHHHHHHHHHHHHHHHHH nnnnnnnm MES 1 2 FERM 10559 2006 3 14 4 FERM 10553 2006 3 14 o oS 6 FERM BP 10555 2006 3 14 8 BP 10557 2006 3 14 3 FERM BP 10560 2006 3 14 ee ei 5 FERM BP 10554 2006 3 14 7 FERM BP 10556 2
21. 10000000 1000000 10000000 EE 000000000 100000 10000 900000000 1004 g 19 0 0000000 100 1000 g kg 1 10u g kg 1 10000000000 1 1 10000000002 30000000000 10000000000000000000 10000 0 0 00 0 IL 3 CSFH neg CSFH 0 SCF LIFO M OSM 0 Ml 10000 1L 1 0 1L 1 IL 20 IL 30 IL 40 1150 1L 60 1L 1100 1 CSF 19 0 MCSFQ 5 0 0 0 0 IFNH HHHHHHHHHHHHHHHHHHHHIFN FN IL 70 IL 80 IL 90 TL 10 1L 12 1L 130 1L 140 1L 15 1L 160 IL 170 11 18 0 Ang 1 20 Ang
22. 1964344110 0000000000 N5KG43440 1000000000 6434 P5 6434 P6 98 0 00 0 100 000000 98 100 55 72011 770000000 98 109 0 68 C 1 30 7292 710 D 00 0 0 O Pyrobest DNA Pol ymerase N5KG3344h1 000 0434 P7 6364 PCR DNA 0 QI Aqui Gel Extraction Kit 000 J j 98 1100 98 10 55 C 30 72 C 1 70 0000000000000 1000000000 0434 P5 6364 PA RO 00000 O 98 10 68 C 1I00000000300 72 7 00 0 PCR 1000000000 Q Aqui ck Gel Extraction Kit 10000000 pcr 4 Bl unt 10000000 DNA 0000000000000 G4344hl0 9 1gG4344uh G4344uh N5KG4344H O00 00000000 00 0 O0 P5 17 1R0 98
23. 1 1 2 Ti PC ISA 210 1 22 2005 4 183 02007 108559 Al 2007 9 27 PCT JP2007 056536 1 13 14 15 17 18 19
24. 15 02007 108559 Al 2007 9 27 100000 10 20 30 40 50 16 02007 108559 1 2007 9 27
25. PC 1 3 2 PC 13 3
26. 9000000 000000 00000 HHHHHHHHHHHHHHHHHHHHHHHHHHHHH RNAH 100000000 LU OU 5 RACE 1000000000000000000000 HHHHHHHIH HHHHHHHHH UU NU 1 1 1 100000000 20060 30 141000000 HHHHHHHHHHHH 142 02007 108559 Al 2007 9 27
27. 10 20 30 40 50 17 02007 108559 Al 2007 9 27 E n E K 5 ee 10 20 30 40 50 02007 108559 1 2007 9 27 18 10 20 30
28. o L3 L3 L3 10000000000000000000000000 00 0 0 100 b 0000 1000000000000 Upper hinge O0 Kabat EU D O O O O
29. 000000 0000000 2 O O FOCP entHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH HHH c MIHHHHHHHHHHHHHHHH DO uz 0000000000000000000000 19610 0 U 7 CFU Mk 100000000 CD34 D HHH HH HH DO 00000000 o D00000000000000 DOO 000000000 DO D DOO 0000 000000 DO D 000000 1964 1904344 196434401 gG4344uh 1964344 0 0 000000000000000 HHHHHHHHHHHH4CHHH4C3HHHHHHHHHHHHHHHN5KG1_7 10 N5KG1_4 490 00000000000000 4D0004D3000000000000000NKGL 6 4 50 N5KG1_6 5 2 00000000000 1000000000000000000000000 HHH4F HHH4F 2HHHHHHHHHHHHHHH 7 10643 44u 00000000000000000 7 10643444 IHHHHHHHHU7TPOHHHHHHHHHHH4 4961 4 49G3311 4 49G3331H D D 0 UT7 TPO 7 10G4344u hm 4 4964344 0 Oooo 7 10G4 344uhm 4 49G4344uhnt HHHHH HHHHHHHH HH HHHHH 6 5 2G 1 6 5 2G3344HHHHHHHHHHHHHHH HH
30. 10 20 30 40 50 20 WO2007 108559 1 2007 9 27 10 20 30 40 50 21 WO2007 108559 1 2007 9 27
31. 1 2 FERM BP 10559 2006 3 14 Anti hMpl 7 10_HV pCR4 4 FERM BP 10553 2006 3 H 14 Anti hMpl 4 49 HV pCR4 6 FERM BP 10555 2006 3 14 Anti hMpl 6 4 50 HV pCR4 8 FERM BP 10557 2006 3 14 Anti hMpl 6 5 2 HV pCR4 3 FERM BP 10560 2006 3 14 Anti hMpl 7 10_LV pCR4 5 FERM BP 10554 2006 3 14 Anti hMpl 4 49_LV pCR4 7 FERM BP 10556 2006 3 14 Anti hMpl 6 4 50 LV pCR4 9 FERM BP 10558 2006 3 H 14 Anti hMpl 6 5 2 LV pCR4 85 86 87 88 89 0 90 92 93 94 0 0 95 96 97 98 99 0 CDR conpl ement ari t y det region 0 000000000000 RASQGI SS T LA 24 341 DASSLESG 00000 0 50 560 0 QO NSYP L O0 WTO 89 970 100000000 10 RASQSVSSSYLA DO 24 35 DASSRAT 51 5700 0 0 OOYGSSPI 1 90 980 PONE es b o Bes H E 10 10000000000000000000000 0000 FMA
32. 10 20 30 40 50 7 02007 108559 Al 2007 9 27 HH 000g 0 HH D D D em ANE 10 20 30 40 50 8 WD2007 108559 1 2007 9 27
33. L1 L1 165 02007 108559 1 2007 9 27
34. GCCAAGGGACCACGGTCACCGTCTCCTCA DO 270 lt 4990 0000000 gt 000000000000000 10000000000000000000000000000000 MELGLSW FLVAI LKGVQCEEQLVESGGGLVQPGRSLRLSCTASGF TF DDYAMY WRQVPGKGLEW SGI 5 5651 GYA DSVKGRFTVSRDNAKNSL QVNSL RAEDTALY Y CAKALWF PHYYGYDVWGQGTTVTVSS O 280 lt 4 491 gt 200000000009 0000000000000 DNA 1000 ATGGACAT GAGGGT CCCCGCT CAGCT CCT GGGGCTT CT GCT GCT CT GGCT CCCAGGT GCCAGAT GT GCCAT CCAGTT GAC CCAGT CT CCAT CCT CCCT GT CT GCAT CT GT AGGAGACAGAGT CACCAT GCCGGGCAAGT CAGGGCAT TAGCAGTA CTTTAGCCT GGT AT CAGCAGAAACCAGGGAAAGCT CCT AAGCT CCT GAT CTAT GAT GCCT CCAGTT T GGAAAGT GGGGT CCAT CAAGGTT CAGCGGCAGT GGAT CT GGGACAGATTT CACT CT CACCAT CAGCAGCCT GCAGCCT GAAGATT TT GCAAC
35. H ipe 10 20 30 40 50 53 02007 108559 1 2007 9 27 HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH omo II EM M E E s nn AE EE o EIE Es HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH 000000 HHHHHHHHHHHHHHHHHhH 7 10VL 3 ATQLTQSPSSLSASVGDRV
36. 10 20 30 40 50 25 02007 108559 1 2007 9 27 H nnnmn X nn 3 10 20 30 40 50
37. 9 0 900 D D lod 0029000000 D D 7 1064344 log 0000000 9B cn 0 9 7 1064344 MIHHHHHH H H 000000 0 00000 000 168 02007 108559 Al 2007 9 27
38. 2 20 000000000000 PCR Bj O C 550000000 5 0000 LM CIO DODO 100000 00000000000000000000 00000 100000 OO 00 000 Q Aqui ck Gel Extraction LM Zero unt TOPO PCR Cl oni n g Kit 0000000000 per 4 Blunt Ph D D B 0 DNAD O D B 0 0 0 M3 20FW 0 M3RV LU hk 5 5 AGG GAG GT 3 20H hh2H 00000 5 GGA GGG GG TCA CCA TG 3 0 0 0 0O 0O 210 1962 1341 5 TGCACGCCGC TGGTCAGGGC
39. 8 1 8 1 e R HHHHHHHHHHHHHHHHHHHHHHHH EP314161 1000000000000000000000000000000 DAT HHHHHHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHH Sequence of proteins of immunological interest Publication No 91 3242 000 0 LH 2288 0000000000000000000000 19540000000 5 50 100000000000000000000 235000000000000000
40. LE DI oo _ m O DI Ba EHE Eee n DI oo DI DI oo DI 2 E oo DI DI DO 3 gt DI cm MES DI i ae Ey EE oo 3 DI Do DI amp See EM DI Do e AES E ED Yes ggo C3 mo EN Lr aS gt US um DI DI L co DI oo DI zm E A l d gt e lt _ NUES i 3 oo oO oO DI era gt DI DI oo C lt CO T u Sii I O Q ET 1 C i 8 oO 1 DI 5 I DI DI 145
41. Mbl0000000000000000000000 WIHHHHHHHHHHHHHc MLHHHHHHHHHHHHH 0000000000000000000000000000000000 IOUUUUUUUUUUUUUUUUUUUUUUUUUUUUUUULU uuu PEG rHuMGDFO OOO 00 Stem 14 651 660 1996 PEG r HUMGDF Dp PR DRS
42. 100 1 0 15m 0 85m 0 0 imag 000 2 4 1 1 10 1 15 1 1 00 00 0000000 05000 3 100000 6900000000000 4 3 DOO0O0O00000000000000000000000 75r0000 5 ni 35m 100 100m 6 100 10000000 10000000 37 CH 5 7 100 10000000 10 8 CD41 100 CFU 00000 100000000 CFU Mx HHHHHHH 7 10 1901 4 49 1960 0000000600 0000 00000000000000000 0000000000000 100000000000 00000000019 L lgk 1 cDNAH DNAD O lgyHHHHHIgkHHHHHHHHHHHHHHHHH 10 5 RACE 5 rapid of cDNA ends
43. 100000 IHHHHHHHHHHHHHH 100 us 4 00000 D D p 10 100 10000 Doo 20 30 55 SEQUENCE LISTING lt 110 gt Kirin Beer Kabushiki Kaisha WO2007 108559 Al 2007 9 27 lt 120 gt Agonistic antibodies to human thrombopoietin receptor lt 130 gt PH 3098 PCT lt 150 gt JP 2006 81322 lt 151 gt 2006 03 23 lt 150 gt JP 2006 299554 lt 151 gt 2006 11 02 lt 160 gt 94 lt 170 gt PatentIn Ver 2 1 lt 210 gt 1 lt 211 gt 163 lt 212 gt PRT lt 213 gt Homo sapiens lt 400 gt 1 Ser Pro Ala Pro Pro Ala Cys Asp Leu Arg Val Leu 1 5 10 Arg Asp Ser His Val Leu His Ser Arg Leu Ser Gln 20 25 His Pro Leu Pro Thr Pro Val Leu Leu Pro Ala Val 35 40 Gly Glu Trp Lys Thr Gln Met Glu Glu Thr Lys Ala 50 55 60 Gly Ala Val Thr Leu Leu Leu Glu Gly Val Met Ala 65 70 75 Leu Gly Pro Thr Cys Leu Ser Ser Leu Leu Gly Gln 85 90 Ser Lys Leu Leu 15 Cys Pro Glu Val 30 Asp Phe Ser Leu 45 Gln Asp Ile Leu Ala Arg Gly Gln 80 Leu Ser Gly Gln 95 10 20 30 40 56 WO2007 108559 Al 2007 9 27 Val Arg Leu Leu Leu Gly Ala Leu Gln Ser Leu Leu Gly Thr Gln Leu 100 105 110 Pro Pro Gln Gly Arg Thr Th
44. h 90000000000000 6400000000000 6400000000 Kabat 100000000000000000000 DNA 10 0 11 uu 2006 813221 0 2006 2995540 0 00 annon HHHHHHHHHHKMIHH HHHHHHHHH
45. n 2u g Ascl Bi o Rad el ectrophoreter 350V 5 CHO 10000 96well culture G418 10000 1000000000000 0 CELL325 PF J RH 2 mM gl ut am 1 00 units m penicillin 100 u g m streptonyci nf hypoxanthine and thym di ne HT 1 100 Invi trogen 5 Mg bsel ect Protein Amer sham Phar maci Bi otech Co Ltd PBSH 20nM Na 50nM NaCl pH3 4 10000000 500 M Ph osphat e Na pH7 0 11 6010 000 1 5 0 Conductivity 4 Ons cm Q Hi trap HPO Amersham Phar maci Bi otec h Co Ltd D SP Sepharose Hi Trap SP Amersham Pharmacia Biotech Co 0 1000000000000 20nM 00 00000000 pH5 0OH 1XPBSH HHHHHHHHHHHHHHHHO0 2 2u ml LLEX GV 10 T B B x HM oanad
46. 010000000000000000000000 utt DO0O0O00007 100000000000000004 400000000 IO00O0O0006 4 50 6 5 20000000000000000000000005 10000 2 35000000000000000000000000000 0000 10 1000000000000 000 10 rnp Protein 0 8x 40cm HHHHHHHHHHH PBSH 0 02M pHaHHHHHHHHHHHHHHHHHHHHHH 1M Tris pH 9 0 pH7 20 0 0 0 0 0 0 0 0 0 0 0 0 00 100 00 Spectrum Laboratories 7 PBSH 0 0 10 224 m0 0000 000 MLLEX GV 0000000 100000 0 0 28017 00 000 0 0 0 1 4 9000000 n E 0 10ng m Recormbi nant Hunan IL 6 R amp D Systens 10 Low IgG Fetal Bovine Serum HyClone eRDFH HHHHHHHHHHHHHHHHHHH 10 5h 97 0 100000 000000000000 10 2 5x10 gt nM 10ng Reconbi nant Human L 6 R amp D Syst ens 1 Low I gG Fetal Bovine 5 HyClone iji 0 O
47. 35 WO2007 108559 Al 2007 9 27 hk 5 5 AGG GAG GCA CCA TTC 3 20 hh2 5 GCT GGA GGG GG TCA CCA CGC TG 3 21 IgG2p 134 5 TGCACGCCGC TGGTCAGGGC GCCTGAGTTC C 3 22 ATGGAGTTGGGACTGAGCTGGATTTTCCTTTTGGCTATTTTAAAAGGTGTCCAGTGTGAAGTGCAGCTGGTG GAGTCTGGGGGAGGCTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTT GATGATTATGCCATGCACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAGTGGGTCTCAGGTATTAGTTGG AATAGTGGTAGCATAGGCTATGCGGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAAC TCCCTGTATCTGCAAATGAACAGTCTGAGAGCTGAGGACACGGCCTTGTATTACTGTGCAAAAAATCTATGG TTCGGGGAGTTCCGTTACTGGTACTTCGATCTCTGGGGCCGTGGCACCCTGGTCACTGTCTCCTCA 23 lU 000000000000000000000000000 HD HB go tt MELGLSWIFLLAILKGVQCEVQLVESGGGLVQPGRSLRLSCAASGFTFDDY AMHWVRQAPGKGLEWVSGI SW NSGSIGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCAKNLWFGEFRYWYFD LWGRGTLVTV SS 24 HHHHHHHH HHHHHHHHHHHHH
48. 3 1201 IgG4PE IgG3311 1gG3331 1gG3344 129334411 1204344 1504344411 IgG4344uh IgG4344uhm 2 35 NT NT NT NT NT NT NT NT NT 7 10 4 49 NT NT NT NT NT 6 4 50 NT NT NT NT NT NT NT NT NT NT NT NT NT NT NT ECs 1 10nM ECs 0 1 1nM ECio 0 01 0 1nM NT 6 5 2 10 20 30 40 50 50 02007 108559 1 2007 9 27
49. 000000 0000000 000000 DNAH HHHHHHHH 4 49 1 7 10 1961 gt HM o N5KG1_4 49 N5KG1 7 10 1 20 0 0 O c 0 0 6 4 50 6 5 0 O0 N5KG1_4 490 0000000000 6 4 50 1960 6 5 2 1961 210000000000000000000000000000 P T Ui v N LVI C 1000000000000 5 00 100000000000001 000000 KOD PI us PCR 5 UJ Di L1
50. 280 0 0 00000 0 01 4 0990000000 UU77TPGdHHHHHHHHHHHHHHHHHHHH 4 49 1gG1 0 9633110 1963330 Al 10 1964344 4 49 1 gG4344uhmj PEG rHuMGDF 0000000000 100000000000000000000000000 7 10 4 490 0 00 0 19610 1gG4PED I gG4344uhm IgG4344uhm 19G4PEH 00 000 19 0000000000000000000 10000000800000000000000000000 3 2 35 7 10 4 49 6 4 50 6 5 2 igG 1 IgG4PE NT NT NT IgG331 1 NT IgG3331 NT NT NT IgG3344 NT NT NT IgG3344h1 NT NT NT NT IgG4344 NT NT NT NT IgG4344h1 NT NT NT NT IgG4344uh NT NT NT NT 1 10nM EC 0 1 1nM EC5o 0 01 0 1nM NT 00000 00000 Epp 000000 000000000000000000 10000 0 1000000 J 5 Ras PI 3K 100000000 1000000000 000000000000000000000000000 1000000000000000000000000000000000000000 1000000 STAT5 Cell Signaling O
51. 40 50 02007 108559 Al 2007 9 27 12 10 20 30 40 50
52. 172 WO2007 108559 Al 2007 9 27 0 HHHHHHHH 000 1000000000 00000000000 ELKTPLGDTTH 0 000 10 CPRCP EPKSCDTPPPCP x3 CPRCPEPKSCDTPPPCPRCP CPRCP 0 VDKRV CH 0000 O APEFLGGPH 0 ESKTPLGDTTHT 11 0000 CH1 0 0 0 J C 2 APEFLGGP 0 l ID JU l l b CD34 CFU MK 10 10 000ng 0 100 b 0000 PEG Tl 0 100 UL
53. 10 HHHHHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHHHHHH P J Del ves ANTI BODY PRODUCTI ON ESSENTI AL TECHNI QUES 1997 W LEY P Shepherd and Dean Monoclonal Anti bodies 2000 OXFORD VERSI TY PRESS J W Godi ng Monoclonal Anti bodi es pri nci pl es and practice 1993 ACAD EM C PRESS
54. nnn0Dn0Onnn0nnnnnnnnnnnnnnnon ra m 1 10 5 40 3H5Sal AGAGAGAGAG GTCGACCACC ATGGAGTTGG GACTGAGCTG GATTT 3 39 3 40 3H3Nhe AGAGAGAGAG GCTAGCTGAG GAGACAGTGA CCAGGGTGCC A 3 40 4 49 5 5 F24HSal AGAGAGAGAGGTCGACCACCATGGAGTTGGGACTGAGCTGGATTT 3 41 3 C15H3Nhe AGAGAGAGAGGCTAGCTGAGGAGACGGTGACCGTGGT 3 42 20 30 40 C C HM e NAVI HOMER ek EM MU nnnnnnnnnnnnnnnnnnnnnn CRIARI nnnnnnnnnnnmn 41 02007 108559 Al 2007 9 27 7 10 LV 5 165 1B_L18Bgl 5 AGAGAGAGAGATCTCTCA
55. g pg qd emu m gg mg g g pmpd pu uu 000 1 10 HHHHHHHH 2 11 100 1000000000000000000 um 100000000000000 lt 40000000000 G4 Kabat 0 0 0 22800 000 000 00 0 235 000000 0 HHHHHHHHHHHHHHHHHHHHH 1 000000000000 0 123 02007 108559 Al 2007 9 27
56. J Y 1 J
57. b UT7 TPO 50 0000000 50 PEG r HuMGDF HHHHHHHHHHHHH 100nM HHHHHHH 1000 Ui DI HH 1961 1962 1963 1954000000000 GenBank Uni Gene n 00228 19620 X03604 K 10000 X64132 X64134 gG4 KO1316 V00557 X64135 X64133H 000 B 1 c 1 1 0 EC50 L1 uu PEG HUMGDF
58. 50 02007 108559 Al 2007 9 27 4 10 20 30 40 100 50
59. 170 02007 108559 Al 2007 9 27
60. 1 r N gt L1 Leukemia 2002 Aug 16 8 1500 1506 HHHHHHUHHHHHHHHHHHHHHHHHHHHHHHHH 100000 KMJ 0 0 HHHHHHH 0000 190 1000001
61. HHHHHHHHHHH E AAR ELE nee HHH HHHHHHHHHHHH 100000 000000000 0 000 00000000 100 Y 100 K Y K 2 ms H s sg S eis sea ERE x RT H 1000000 CHH 0000 a OERE 5 5 PRA MD a E AE 3 zi 00 0 0 L1 34 WO2007 108559 Al 2007 9 27
62. 40 50 1000 C 19 02007 108559 1 2007 9 27 0 1000000000000 0 1000000000000 1000000000000 1000000000000
63. BER TEE EIL TTACTGTCAGCAGTATGGTAGCTCACCGAT CACCTTCGGCCAAGGGACACGACTGGAGATTAAACGIT 37 100000 lt 6 5 200000000x 0000000000000090 10000000000000000000000000000000 METPAQLLFLLLLWLPDTTGEI VLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWY QQKPGQAPRLLI YDASSRATGI P DRFSGSGSGTDFTLTI SRLEPEDFAVYYCQQYGSSPI TFGOGTRLEI 380 00001 1000000 UH 10000000000000000 IOUUUULU HHHHHHHHHHHHHHH N5KG1 Lark N5KG1 0 0 O O DEC Phar maceuti s O US patent 60013581 HHHHHHHHHHHHHHHIH N5KG1 00000000000 wood HHHHHHHHHHHHHHHIH 1900000 IlHHHHHHYHHHHHHHHH HHHHHHHHHH 10000000000000000
64. PEG NH SPAPPACDLRVLSKLLRDSHVLHSRLSQCPEVHPLPTPVLLPAVDFSLGEWKTOMEETKAQDI LGAVTLLLEG VMAARGQLCPT CLSSLLGQLSGQVRLLLGAL QSLLGT QLPPQGRTTAHKDPNAI FLSFOHILLRGKVRFLMLVGGSTLCVR RAPPTTAVPS COOH PEG rHuMGDF 50 0000000 50000000 100nM 0000 nn 10000000000000000 000000000000 000 a O b a 0000 CD40 000000 0000000000000 1 000 0 LU UU UU LU 5 UT7 TPO 1000000000000 0 000 PEGrHuMGDF 70 EC50 10nM 1000000000000000000 b a CD34H 000000 0000000000000 10019 nt 00 0 U 5 UT7 TPO PEGrHuMGDF 90
65. H Y 10 20 30 40 50 51 02007 108559 1 2007 9 27 E 006000000 C C n nm n 10 20 30 40 50 52 02007 108559 Al 2007 9 27
66. 26 JP 02007 108559 1 2007 9 27 B SN B ni 0 DO 10 20 30 40 50 27 02007 108559 Al 2007 9 27 C 4 ART m C 1_ 1 5 AGAGAGAGAG GAATTCGCCA CCATGCCCTC CTGGGCCCTC TT 3 12 Mp1_R2 5 AGAGAGAGAG CGGCCGCTCA AGGCTGCTGC
67. VI 7 10G4344uhm 10 gf 10 TPO HHHHHHHHHHHHTPO 000 0 0 00000 7 10G43344uhm 7 10 7 10VL 4 49 1040 0000 0 0 6 4 500 00 0000000000000 0 43 61000 7 10G4344uhn O00 0000000000000000 7 10G4344uhm CORO 4 HHHHHHHHHHHH 0 100 7 10VL HHHHHHHHHHHH lt n 7 10VL Al QLTQSPSSLSASVGDRVTI TCRASQG SSALAW QOKPGKAPKLLI YDASSLESGVPSRF SGSGSGTDFTLTI SSLOP EDFATYYCQQF NSYPLTF GGGTKVEI 7 10VL_V104L 4 49VL 0 850 Al QLTQSPSSLSASVGDRVTI TCRASQGI SSALAW QOKPGKAPKLLI YDASSLESGVPSRF SGSGSGTDFTLTI SSLOP EDFATYYCQQF NSYPLTF GGGTKLEI K 7 10VL_G100Q 6 4 50VLH 2 0 U 0 8600 Al QLTQSPSSLSASVGDRVT I TCRASQG SSALAW QOKPGKAPKLLI YDASSLESGVPSRF SGSGSGT DFTLTI 551 EDF ATYY CQQF NSYPLTF GQGTKVEI 7 10VL A43V 6 4 50 2 0 000 870 0 Al QLTQSPSSLSAS
68. kali ne Phosphatase E coli C75 000000 8 9 DNAU 0000000020 00 0 0 lig ase Lt l DNAD DNAD LVO N5KGl 000 5 000000000000000000000 N5KG1_6 4 50 Lv N5KG1 6 5 2 Lv 000 Ul 0000000 000000000 HV C H 0 0 0000 DNA UL 5 Sall 3 Nhel UL 6 4 500 5 50 5 7Hsal 5 AGAGAGAGAG GTCGACCACC ATGGAATTGG GACTGAGCTG GATTTT 3 7 0000 510 3 C15H3Nne 5 AGAGAGAGAGGCTAGCTGAGGAGACGGTGACCGTGGT 3 520 6 5 20 5 000000 0 24 5 3 0 0 0 530 3 00000 0 0 L66H3Nhe 5 AGAGAGAGAGGCTAGCTGAGGAGACGGTGACCGTGGTC 3 0 540 94 1010000000 94 50 68 45 35 72 C 70
69. TTATTACT GT CAACAGTTTAATAGT TACCCGTACACT TTT GGCCAGGGGACCAAGCT 290 4 49 2 0000000000000 HHHHHHHHHHHHHHHHHHHHHHHH MDMRVPAQLLGLLLLWLPGARCAI QLTQSPSSLSASVGDRVTI TCRASQGI SSTLAW QOKPGKAPKLLI YDASSLESGV PSRFSGSGSGTDFTLTI SSLQPEDFATYYCQOF NSYPYTFGOGTKLEI KR 30 1000000 6 4 500 0000000000000 DNAHHHHH JH 0191001001100 1000 lt 6 4 500 0 00 0 0 gt 20 00000000004 DNA ATGGAATTGGGACTGAGCTGGATTTTCCTTTTGGCTATTTTAAAAGGT GT CCAGTGT GAAGT GCAGCT GGI GGAGT CTGG GGGAGGCTTGGTACAGCCT GGCAGGT CCCT GAGACT CT CCT GT GCAACCT TCACCTTTGATAATTATGCCATGT ACT GGGT CCGGCAAGCT CCAGGGAAGGGCCT GGAGT GGGT CT CAGGTAT T AGT T GGAATAGT GGT GACATAGGCT AT GCG GACT CT GT GAAGGGCCGATT CACCAT CT CCAGAGACAACGCCAAGAACT CCCT GTAT CT GCAAAT GAACAGT CT GAGAGC TGAGGACACGGCCTT GTATT ACT GT GCAAGGGAT GCGGGGT T CGGGGAGT TCCACT ACGGT CT GGACGT CT GGGGCCAAG GGACCACGGTCACCGTCTCCTCA DODO 100000 lt 4 5000000000 gt 2000000000000000 00000000000000000000000000000 MELGLSW FLLAI LKGVQCEVQLVESGGGLVQPGRSLRLSCAT SGF TF DNYAMY WRQAP GKGLEW SGI SWNSGDI GYADSVKGRF TI SRDNAKNSLYLOMNSL RAEDTAL YY CARDAG FGEFHYGLDVWGQGTTVTVSS 0 32 100000 6 4 500 10000 ATGGACAT GAGGGT CCCCGCT CAGC
70. gt ATG TTATTACT GT CAACAGTTTAATAGT TACCCGCT CACTT T CGGCGGAGGGACCAAGGT O 25 lt 7 10000 0 20 0 2 0 000 0 0 0 00 0 0 0 10000000000000000000000000000000 MDVRVPAQLLGLLLLWLPGARCAI QLTQSPSSLSASVGDRVTI TCRASQGI SSALAW QOKPGKAP KLLI YDASSLESGV PSRFSGSGSGTDFTLTI SSLQPEDFATYYCQQF NSYPLTFGGGTKVEI 260 155 02007 108559 Al 2007 9 27 100000 4 49 00 000000000000 0000000 DNAD O 000000000000000000000000000000000 4 497 000 gt ATG 100000 1000 ATGGAGTT GGGACT GAGCT GGATT TT CCTT GI GGCTATTT TAAAAGGT GT CCAGT GT GAAGAGCAGCT GGT GGAGT CT GG GGGAGGCTT GGT ACAGCCT GGCAGGT CCCT GAGACT CT CCT GTACAGCCT CT GGATT CACCTT T GAT GATT AT GCCATGT ACT GGGT CCGGCAAGT T CCAGGGAAGGGCCT GGAGT GGGT CT CAGGTAT T AGT T GGAACAGT GGT AGCATAGGCT AT GCG GACT CT GT GAAGGGCCGATT CACCGTT T CCAGAGACAACGCCAAGAACT CCCT GTAT CT GCAAAT GAACAGT CT GAGAGC TGAGGACACGGCCTTATATT ACT GT GCAAAAGCCCTAT GGT T CGGGGAGTT CCCCCACT ACT ACGGTAT GGACGT CT GGG
71. U 0 0 00 0000 0 0 0000 00 HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH DO0O00000000000000000000000000 Arg0 Hs L 5 0000 00 00 Leur Val GyH Ser Asng Gn H Phell Tyro Trp HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHHHHHHH NCBI HH D00000 0 0 0 FASTAHHHHHHHHHHHHHHHHHHHHH 143 02007 108559 Al 2007 9 27 1000 1000000000000000000000000 90000000000 PCRHHHHHHHHHHHHHHHHHHHHHHHHHHHH 1000000000000000000000 Sanbrookg Molecular Cloning boratory Mannual Cold Spring Harbor Laboratory Press 1989
72. 100 LU Cb CD34 100 9 10 b UT7 TPO PEGrHuMGDF 90 d EC50 1nM 100 1000000000000 6 00000 00000 122 02007 108559 Al 2007 9 27
73. TPO MO J ak STAT PI 3K Akt Ras 0 O c 10 200 TPO ccMIH HH 1000000 10000000 0 0 00 00000000 000000000000000 00000 00 00000000 000000000000000 0000000 000000000000 000000000 0 100000000000000000000 TPO HHHHHHHHH 1000000 rhTPQ 00 0 O TPO 0 163 00 PEG PEG HuMGDF ated Reconpinant Hunan Megaka yocyte Growth and Devel opment Factor E I di opat hic Thrombocytopeni c Purpura ITPH 100000000000000000000000000000000000000 Aplastic Mel odyspl asti c Syndrom MSU 00000000000 10000000000
74. lt 6 5 21 00000 gt 000000000090000000000000 DNAD ATGGAGTT GGGACT GAGCT GGATT TT CCT TT TGGCTATTT TAAAAGGT GT CCAGT GT GAAGT GCAACT GGT GGAGT GT GG GGGAGGCTT GGT ACAGCCT GGCAGGT GAGACT CT CCT GT GCAGCCT CT GGATT CACCT TT GATGATT AT GCCAT GC ACT GGGT CCGGCAAGCT CCAGGGAAGGGCCT GGAGT GGGT CT CAGGTATT AGT TGGAATAGT GGTAGTATAGGT TAT GCG GACT CT GT GAAGGGCCGATT CACCAT CT CCAGAGACAACGCCAAGAACT CCCT GTAT CT GCAAAT GAACAGT CT GAGAGC TGAGGACACGGCCTT GTATT ACT GT GCAAAACCT ATAT GGT T CGGGGAGT GGGGAAACT ACT ACGGT AT GGACGT CT GGG GCCAAGGGACCACGGTCACCGTCTCCTCA 0 0 350 1100000 d ge E B 10000000000000000000000000000000 MELGLSW FLLAI LKGVQCEVQL VECGGGL VOPGRSL RLSCAASGF TF DDYAVHW RQAPGKGL EW SGI SWNSGSI GYA DSVKGRFTI SRDNAKNSLYLQMNSLRAEDTALYYCAKPI WFGEWGNYYGMDVWGQGTTVTVSS 0 0 DD 360 1100000 lt 6 5 21 00000 20 ATG 1000 ATGGAAACCCCAGCGCAGCTT CT CTT CCT CCT GCT ACT CT GGCT CCCAGATACCACCGGAGAAATT GT GT T GACGCAGT C TCCAGGCACCCT GT CTTT GT CT CCAGGGGAAAGAGCCACCCT CT CCT GCAGGGCCAGT GT TAGCAGCAGCT ACT TAGCCT GGTACCAGCAGAAACCT GGCCAGGCT CCCAGGCT CCT CAT CTAT GAT GCAT CCAGCAGGGCCACT GGCAT CCCA GACAGGT T CAGT GGCAGT GGGT CT GGGACAGACT T CACT CT CACCAT CAGCAGACT GGAGCCT GAAGATT TT GCAGT GTA
75. DO00000000000000000000000000000000 HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH 1000000000000000000000000000000000 HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH 30000000000 1000000 0 100000000000000 HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH 8 BALB c P3X63Ag8U 10 P3 Yel ton DE et al Current Topics Mcrobiol and Im unol 81 1 70 1978 P3 NSI 1 Ag4 1 NS 1 Kohler et al European J Immunology 6 511 519 19760 Sp2 O Ag14 SP 20 Shul M et al Nat ure 276 269 270 1978 P3X63Ag8 6530 653 Kearney J et al J nology 123 1548 15509 19791 P3X63Ag8 X630 Hori bata K and Harris W Nature 256 495 497 19759 0 10000000000 100000000 38 0000 1000 1002 0 0000000000000 HHHHHHHHHHHHH FCS 0 16401 0 0 8 0000 10000000000000 1000 15 s Modified becco s Med ium O0 0 0 0 1MM 0000000000 0 Dul 5 Modi fi ed Eagle Medi O00 00 DVEMI 00000 1000030 40000 100 0 0 000000000000 00 2 10700 00 4 0000 00000 DMEM 0 0 0 0 0 0 000000000000000000000000 5 10 10 1 0000000 img 0
76. 10 20 30 40 50 31 WO2007 108559 1 2007 9 27 0 H 7 0 0 0000y uU H H UH C D 10 20 30 40 50 32 02007 108559 Al 2007 9 27 100000000000000
77. sterile HO 28u cDNA 2 5 1 KOD PI us buffer 10X dNTP M x 2n M 5 MSO 25 mM 21 KOD us 1 unit ul Universal primer A mx UPM 10X Gene specific primers GSP 101 1 5 Total volume 50 100 1000000000000 SMART RACE cDNA Anplification Kit 0 0 0000 1961 00000000000000000000004 00000 hk 20 0000 1gG1p 00000 5 TCTTGTCCACCTTGGTGTTGCTGGGCTTGTG 3 18H
78. 2 001 00 37 3 10 cel s nt 000 1000000 50u 96 well plate 4 00000000000000 1940000000000 494000000 50u 100000000 5 37 5 O 00000000000 6 8 10000000000 10000020000000 7 TECAN SUNRISE RAINBOW 100 0 0 450m 0000 eoonn 7 100 0 44911 O 6 4 5009 00 6 5 2000 000000 UT7 TPO c MIH UT7TP 0 50 EC5OH MexX 2 UTI TPO EC50 UT7 TPO Max UT 2 35 1 uri _m _ _ _ 7 10 IgGi gt 90 1 4 49 IgG1 gt 80 2 6 4 50 IgG1 gt 80 3 6 5 2 161 gt 50 3 PEG rHuMGDF 0 001 0 01nM 100 MHR ECso 1 1 0nM
79. colony count 000 0 GM ony f or m ng uni t granul ocyte macr ophage erythroi 00 meant 50 0 Vehi lt 100000 0000 0 PBS NT Non treat DOO CD34 41 1 00000000 10 0001 Ul 1000000000000 Vehi cle PBSH NT Non treat ed uu Mel 1000000000000000000000 Tg 1000 PBS H TgH 00 Non T9 7 10G4344uhm 101 90 ER ph F P 5 7 10G4344uhn 0000000 000 20 7 10643440 000000 00 U 7 TPO l 11 UH2G3uhm 120 16 180 22 390 54 utu 57 73 74 G3344h1 751 G3344 761 64344 77 0434411 780 G4344uh 79 G4344uhm 800 811 7 10G4344uhm 820 7 10G4344uhm 830 7 10G4344uhm 840 7 10G4344uhm 851 7 10VL V104L 860 7 10VL_G100Q 87 7 1 _ 4 D H 21 3 9 2009 3 9 O 10000000 10000 100 2 0 1 HHHHHHHHHHHHHHHHHHH 2
80. 1 ITPH 2 0 3 4 MDS 5 0000 174 WO2007 108559 Al 2007 9 27 6 0000000000000000000 HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH u a ELA aa 175 02007 108559 Al 2007 9 27 00000000 INTERNATIONAL SEARCH International application No PCT JP2007 056536 A CLASSIFICATION OF SUBJECT MATTER C12N15 09 2006 01 1 61 39 395 2006 01 1 A61P7 00 2006 01 i A61P7 04 2006 01 1 A67P43 00 2006 01 i C07K16 28 2006 01 i CO7K16 46 2006 01 i C12N15 02 2006 01 i 12 21 08 2006 01 1 According to International Patent Classification IPC or to both national classification and IPC FIELDS SEARCHED Minimum documentation searched classification system followed by classification symbols C12N15 00 15 90 C07K16 00 16 46 Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched Electronic data base consulted during the international search name of data base and where pr
81. 02007 108559 Al 2007 9 27 5 10 20 30 40 50 02007 108559 Al 2007 9 27
82. 08 06 2007 RIO TRIO BETETE BAER I S AZ J P GRG 100 8915 43 03 3581 1101 3448 1 210 2 20054445 181 02007 108559 Al 2007 9 27 PCT JP2007 056536 C TA 2004 11 13 211 7 733 736 JP 2002 12776A 2002 05 08 WO 1999 055369 Al amp 1073464 A1 amp US 2002 0062009 Al amp 1073464 1 amp DE 69920897 amp ES 2230848 amp US 6936698 82 amp US 2005 0208625 Al amp DE 69920897 2 WO 1999 010494 A2 GENENTECH INC 1999 03 04 amp AU 1998 088312 amp 1009831 2 amp JP 2001 513999 A amp US 6342220 1 amp AU 755822 amp AU 2003 201322 1
83. 140000000000 LE ERETTE 7 Mol 2 S Mp1H I 0 100000 0 HI VO TPO 100000 Wg ED 12 00 0000 Mo i B m B B n Q n ET D B DO nl TD C7E3 xi mab ENE MB n D EE DE 02007 108559 Al 2007 9 27 144
84. 6 4 50 5500000 208LF AGAGAGAGAGAT CT CT CACCAT GGACATGAGGGT CCCCGCTCAGC 3 47 3 00000 62LP3Bsi AGAGAGAGAGCGTACGTTTGATTTCCACCTTGGTCCCTTG 3 6 5 2 rur 48 5 0000000 27 AGAGAGAGAGATCTCTCACCATGGAAACCCCAGCGCAGCTTCTCTTC 3 00000 20218 3 0 0 0 D D 500 1000000 94 CH 1 68 450 35 72 C 7 Bgl Il 5 400521 5KG1H 000000000000 110 Bsi W 100000 490 L3 u r 160 WO2007 108559 Al 2007 9 27
85. 1 000 ng O LU 100000000000 b 7 0000000000000 od 0 PEGrHuMoor 70 000 10nM LU iii 000 60000000000 000000000000 0 CD34 100 ng nt LU 0 b UT7 TPO HH PEGrHuMGDF 90 0 EC500 1nM 100000000000000000000000000000000 HHHHHHHHHH HHHHHHHHHHHHHHHUHHHHHHHHHHHHHHHHHH 000000000000000000 7 109 130 02007 108559 Al 2007 9 27 b D 100000000000000000 000000 4 49 g abr BB HHHHHHHHHHHHHHHHHHH6 4 50H D D HH D 6 5 20 100000000000000000
86. 0 0 G 000 1L 3RPU c MolH 0000 CRH Cytoki ne Receptor Honol ogue CRH1 CRH20 0 WV5XV6 1 2 uu 0 0 0 0 Mol Coen
87. 0000 1 1 0000000 00 100 100000000000000000000 000000000000000000000000 000 000000000000000000 000 100 FDCP2H Mol FDCP hMpl FEBS Lett 1996 Oct 21 395 2 3 228 34 0 LU 10000000 0000 10000000000 100 utu uut MHC 0000000000000 100 00 hMlt 0 1020 00000000 19290 0 FMBA 0 PHU 00000 0 5080 0 Trp Ser 000000000 0 Abe Leukemia 2002 Aug 16 8 1500 1506 LD HHHHHHHHHHHHHHHHHHHHH 000 00 hMol Ser H 1 000 c Molp hMol 0000 hMpl cDNAI Pas12 Bartl ey Cell 199 4 Jul 1 77 7 1117 1124 Morita H FEBS Lett 1996 Oct 21 395 2 3 228 234 PCRH l 100 105 3 Xbal Mol F1 Mol R2 0 0 O KOD Plus DNAD uu PCRI PCR O0 O0 O 97000 LU
88. 5 AGAGAGAGAGATCT CTCACCAT GGACAT GAGGGT CCCCGCT C 3 0000 430 159 02007 108559 Al 2007 9 27 10300000 165 18 118 CGTACGTTTG TGGTCCCTCC 3 O 440 B 5 0000000 1189 p AGAGAGAGAGATCTCTCACCATGAGGGTCCCCGCTCAGCTC 3 0 0 0 0 D 450 3 27 02 3 0 D 0 D 0 460 ur UJ 94 1 94 C 5 68 C 450 72 7 0000 L 0 4BI unt TOPOH 000000000000000 DNA DNA DNA ODO ML3 20FW M3RV 0 0 0 2 7 10 TOPO 4 49 Lv BlIIH Bsi W 00 400 bp DNA Bgl Il Bsi W 7 10 4 49 000 9 3kb T4 DNA ligase 1000000 DNAD
89. 0 100000000000000000000 10 10 0 24 00 0000000000000 nnm 0000 bi 0 10000000000000000000000000 hMpl Mol O FDCP2 FDCP hMpl 0 FEBS Lett 1996 Oct 21 395 2 3 228 34 000 n 4 10 0 0 50 u LO FACS stai ni ng medi 2 FBS 0 1 NaN lnM E DTA in PBS SO0ULHHHHHHHHHHHHHHHHHHHHHHHHHH 0 1 14 9000000000 300 0000000 FACS staining medi 0000 R phycoerythrin 0 ab 0 South ern otechnol ogy Cat 2043 0908 60000000 0 300 00000000000 Propi di I odi de O FACS stai ni ng medi um 000000 FACS Becton Di 10000000000 000000000000000 0 0 0 FOP h 00000000000000 FDCP20 0 FDCP parent 100000 vl 0000000000000 0000 97 1 00000 0000000000000 1000000000000 00 0 1177 000000000060 000000 7 TPQ aki et al Blood 1998 Dec 15 92 12 4652 62 GDF O Iscove s Modified Dulbecco s Medi um MM 000000000000 151 02007 108559 Al
90. pj 30 40 50 02007 108559 Al 2007 9 27 3 10 20 30 40 1000
91. Nat Bi ot echnol 1996 Vol 14 8450000000 E th b D PB FB DB D D Ep Hod 19910000 1 00 1000 lt 0 000000000 0 929 hMl hol c MID i FMBA Ser 0000000000000000000 149 02007 108559 1 2007 9 27
92. 0 0 0 DMO unt 00000000 DNAD DNA M3 20FW DN DNA 6 4 50 Hv TOPO 6 5 2 Hv DNA 0 O O salig Nnel OO 430 DNA SallH OOO 6 4 50 6 5 20 10000000000 9 3kb D D 0 0 0 0 0 DHOB 10000000000 DNAD 0 DNAH Od 000 10000000000000 100 6 4 50 IgGl 6 5 2 1961 00000 N5KG1 6 4 50 N5KG1 6 5 2 000 N5KG1 6 4 50 N5KG1 6 5 20 000 00000 20 9 ce Melr B D 00 N5bKG4PEH NbKG4 PE 0000 Bang c MID 0 0 O 5 1 7 100 4 49 N5KG4PE 7 100 NSKG4PE_4 49 00000 3 N5KG3 gG3 10000 196 1 00000000000 196300000 UL 19630
93. HHHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHHH 0000 Faby 1000 0000000000000 L1 134 02007 108559 Al 2007 9 27 0000 0000 HHHHHHHHHHHHHHHHHHH 0000000000 00000 00000 EpoR G lt 0000
94. 10000000000000002000000000000000 p 441 1 0000 10 00000 0 0 0 0 0 0 0 0 00 0 000 167 02007 108559 Al 2007 9 27
95. 5 7 10 Gl 4 49 5 6 4 50 5 1 6 5 2 HHHHHHHHHHHH 7 10 IgG4344uhn 0 DOO 7 10_1gG4344uhr DOO 00001 000c Mbl 293F DNA EndoFree Plasmd Kit FreeStyl eTM 293 Expression Syst em nnnnn nn 280 000000000 E 500 g Hi Trap rProtein PBS 10 20 nM pH7 0 B N 1000000000000000000000000000000 dhf r CHO DG44 I DEC Pharnaceuti cal s Corporati on EX CELL325PF J RO 0 0 10000000000000000000
96. 1 2 1180368 Gi 1 3 118 0368 4 1 4 A OFA Gv 4 1 5 1 6 41803888 8 1 7 8 v 8 8 8 vii 1 9 2 0 1 2 7 PC 1 SA 210 2005 4 184 02007 108559 Al 2007 9 27 1000000000 5I nt d 0000 2708 2006 01 0000 2 08 80000 AP LS NA SD SL SZ TZ UG ZM ZW EN AM AZ BY KG KZ M RJ TJ EP BE BG CH CZ CK ES Fl FR HJ I E I S IT LT LU LV MC M PL PT SE 9 SK ON BF B G GN Q M NE SN TD TG AG AL AMAT AU AZ BG BR BWBY EZ NORM CZ LE DK EM LZ EG ES FI GM GI FN FR HJ ID IL IN LS J P KE KG KP KZ
97. 4B CHL CH2 VDKRV CPPCP APELLGGP IgG2 VDKTV ERK CCVECPPCP APPVAGP IgG3 VDKRV ELKTPLGDTTHT CPRCP EPKSCDTPPPCPRCP x3 APELLGGP 1264 VDKRV ESKYGPP CPSCP APEFLGGP G4PE VDKRV ESKYGPP CPPCP APEFEGGP G4344 BLKTPLGDTTHT CPRCP EPKSCDTPPPCPRCP x3 APEFEGGP 043441 ELKTPLGDITHI CPRCPEPKSCDTPPPCPRCP APEFEGGP G4344uh VDKRV ELKTPLGDTTHT APEFEGGP G4344uhm VDKRV ESKTPLGDTTHT CPPCP APEFEGGP Bl4c 1 Pacl ASTI D ane ORS gt n 3 Il 1 ma Jv 5 5 v UE 8 3 E E E ns 8 4 aw n 42 gt x x 02007 108559 Al 2007 9 27 114 ues 18UN NON e PID BIGc ozud u oco6b b 01 XXX XXX 0401 1854 RON 612 Ped t l forze BAY SEGA GHE are 27665 z infe YadqoorGs 169 14993 148889556 HG 0 0d0 1 1unigp 429 Id 1953 tasy H 2 0 6 amp amp Eu EE usd Iso h 8 HRAL 665 nes 84 as 194N
98. 2005 056604 128 02007 108559 1 2007 9 27 5 6 1 HHHHHHHHHHHHHHHHHH 11 Ut HHHHHHHHHHHHHHHHHHIH HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH a 0 000000 OMM 0000000000000 10 000ng nt
99. 5 10 10000000 900000000000000000000000 5x 10 DOO0O0O0000030000000300000FMBAPhMlODOD 2x 1060 0o00 5u 99 000000000000000000 100000000000000000100000 00 000 10u 000000000000 00000 x 0000 0000 Se 900 l 1929 5x 1400000000000000000000000O 000 000 00000 5p 90 00 0 10 60 5u 99000000 000 00000000 li l 0 000000000000000 350ng nm l 50 m 50 g m D 10m0 0 000 1000000000000000000 LU LU 0 DVEM 2000000000 D M LU 10 FCS DMEM 3720 50 10000000 1 108 ml SP2 ATCC CRL 1581 DMEM DMEM 000000 5 1 OCA EE 500 wv 15000000000 0 lm 0 37 DVEM 1m 2000 000 7 DME 0 000 LU 10 Fe Calf Serum FCS 00 LU DMEM 100 FCS
100. MbTrap GII 00 0 139 WO2007 108559 Al 2007 9 27 1000000000000000 10000 Ouchterl ELI SA RIAODOODOOOOOOOOOOOOO0OOO0OO0OO0OO0O0O00000000000 ELI SAD 0 0 D 0 0 0 0 0 0 0000000000000 0000000 000000000 D0O000000 0000000000000 28017 U 0 0 0 0 D 1 40 0022800 000 DD D D 00 ing mHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH
101. Sequences of Proteins of Inmunological Interest 5 Ed Public Health Serv ice National Institute of Health Bethesda 1991 10 21e 000 2 26 N Mddle 0000 0 22600 lJ 2310 DOO0O0O0O000000000c0000000000000 OOOO 000000 uu CH2H 000000 00 00 lover hing 1000000000 amp 00000000000000 0 Kabat 22800 0 0 0 0 0000 0 0 2350 0 1000000000000000000000 100 0 000000000000000000 g 131 02007 108559 Al 2007 9 27 9 uuu G 00000 6960000000000 rt 2281 0000000000 0 2350 00 0000000000000 L1 rn Co Co 0 11 11 a h 1
102. uum HHHHHH HHHHHHHHHHHHHuHH Hu 100000 HHHHHHHHHHHHHHHHHH HHHHHHHHH 000 100000000000000000000000000000000000 10000000000000000 00000000000000000 IgGlp 7 5 TCTTGTCCACCTTGGTGTTGCTGGGCTTGTG 3 18 hk 2 5 GTT GAA GCT TGT GAC GGG CGA GC 3 19 CH un 0 c0 goig 100000000 1000000000000 H D 100000 1000000000000 1000000000 ENTE Do H H 000 H H H H 00000 000 L H H H H nnn nn nnnnn nn nnn nnnnn nnnm CH n n 00000 0000
103. 7 10HV 4 49M 0o 0 0000000 0000000000000 7 100 5 40 3H5Sa 5 AGAGAGAGAG GTCGACCACC ATGGAGTTGG GACTGAGCTG GATTT 3 0 0 0 0 0 390 3 40 3H3Nhe 5 AGAGAGAGAG GCTAGCTGAG GAGACAGTGA CCAGGGTGCC 3 40H 4 49 5 F24HSal 5 AGAGAGAGAGGTCGACCACCATGGAGTTGGGACTGAGCTGGATTT 3 41H 3 C15H3Nne 5 AGAGAGAGAGGCTAGCTGAGGAGACGGTGACCGTGGT 3 42H 100000000 94 110000000 94 C 5Q 68 C 451 103590000000 72 7 1000000000 0241 000000 Sall 0000000000000 430bp DNAD N5 KG1 SallH eip HHHHHHHAk aline Phosphatase coli 75 10000000000 10000 OO O00 0 8 9 DNAH OOO 0002 4 110 DHIOB O0 000000000000000 DNAHHHHHDNAHHHHHHHHHHHHH 5 10000 DNA N5KG1 7 10 5 1 4 49 uuu 1000000000000000001 00000000000 10000000 1 4 amp 10000000000 0000000000 100005 103 BsiWH 7 100 L 550000000 165 18 11889
104. 10000 1000000000 0 0 0 DNA 1000000000 0 LO 0 MI H B l Hi 00000000 DNA PCR 1000000000 DNA 000000000000000000000 0000000 000000000000000000000000000000 DNAHHHHHHHHHHHHHHHHHHHHHHHHH 000 100000000000000000000000000000000 000000000000000000000000000000 DNAH AHHHHHHHHHHHH 1000000 HHHHHHHHH 000 COSY 1000000 N5KG1 Lark I DEC pharmaceuticals US patent 6001358 000 CW 000 K ND 5 nn K SET nn E CDM gGl N5KG4PE 0 O I DEC pharnaceuti s 2000000000000 Ser 228Pro Leu235G u 140 02007 108559 1 2007 9 27
105. Antibody dependent cellular cytotoxicity ADCO o 0 0 0 0 0 0 0 0 0 0 0 00 00 00 0000 0 19640 IgG4PEH 0 1000 DO0O000000000000000000000000000000000000 02007 108559 1 2007 9 27 1063 11960 000000000
106. 0 BD SMART RACE cDNA Anplification 100000000000000000000000000000 cDNA 1000000000000 7 10 4 49 6 4 50 6 5 20 Total RN total 15 99000000000 Ist strand cDNAU lst strand cDNA Total RNA lH 9 5 CDS 11 SMART Oligo 1 5x Buffer 2h DIT lu DNTP mx 11 PowerScri pt Reverse Transcriptase 1 10 0 4220 1 5000000000000 0 00 506 1 Tricine EDTA Buffer 00 0 0 0 72200 0 0 0 0 0000 00 0 1 51 strand cDNAI 153 WO2007 108559 Al 2007 9 27 20 PCR IU U 2 1 PCR ug 3 00000 BD SMART RACE cDNA cation Kit 0D 0 0 0 cDNA 5 i 0 55000000 Universal pri ne A mx 0 PCR PCR 0 KOD Plus DNAD 000 10000
107. 157 02007 108559 1 2007 9 27
108. 0000 000000000000000000 10000 10000 ES 10000 100000000 100000000 100000000 1000000000000 100000000000 O Wight G et al 1991 Bi o Technol ogy 9 830 834 100 000000000000000000000000000001 1000001000000000000000000000000000 100000000 141 02007 108559 Al 2007 9 27 100000 HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH d 000000000000000000000000 DNA 100 1000000000 100 h 10000000000000000000 DNA s
109. 100 b UT7 TPO PEG NH SPAPPACDLRVLSKLLRDSHVLHSRLSQCPEVHPLPTPVLLPAVDFSLGEWKTOMEETKAQDI LGAVTLLLEG VMAARGQLCPT CLSSLLGQLSGQVRLLLGAL QSLLGT QLPPQGRTTAHKDPNAI FLSFOHILLRGKVRFLMLVGGSTLCVR RAPPTTAVPS COOH PEG r 50 0000000 500 00 0 0 1007 100 c Mol H l 00 0 CD34 1 ooong nt b 70 000 EC50 10nM
110. 1000 PEG NH SPAPPACDLRVLSKLLRDSHVLHSRLSQCPEVHPLPTPVLLPAVDF 5 LGAVTLLLEG VMAARGQLCPT CLSSLLGQLSGQVRLLLGAL QSLLGT QLPPQGRTTAHKDPNAI FLSFQHLLRGKVRFLM VGGSTLCVR RAPPTTAVPS COOH PEGrHuMGDF 50 1500 EC500 100 1 HHHHHHHHHHHHHHHHHH D b 124 WO2007 108559 Al 2007 9 27
111. 0 0 N5KG4PE H 17m 220 G3G4 RI D D D PCR DNAI QAquick Gel Extrac tion Kit 1000000 54000000000 DNAD O 10000 94 C 309 55 O O 72 C 1 164 02007 108559 Al 2007 9 27 4 C 7 100000 qui Gel Extraction Kit 94 C 30 68 C 1 3 72 PCRH 1000000 pCR 4 Blunt 0000000 DNAHHHHHHHHHD u 19 4344 0000000000000 17m 180 5 TGTGTGAGTT GTGTCACCAA GTGGGGTTTT GGACTCAACT CTCTTGTCCA CCTTGGT 3 72 17m 2 5 ACCCCACTTG GTGACACAAC TCACACATGC CCACCATGCC CAGCACCTGA GTTCGAG 3 73 11 HHHHHHHHH HHHHHHHHH DNA 00 Nhel Banti 00000 i
112. 00 000000000 1 V 100000 0000 10 CDR grafting 100 10 00000 Diabody gt sc FV gt 0000 100000000000000000000 100000000000000000000 100000000000000000000 10000000000000000000 a oa a L3 oa L3 L3 oa oa a Japanese Journal of Trans ine 46 3 311 316 2000 MJ Stem Cell 16 1 6 1998 Kuter Blood 100 10 3457 69 2002 Yonenura Int Henat 82 307 309 2005 Komatsu Nj Blood 96 296a 2000 Broudy Cytokine 25 2 52 60 2004 Wang Clin Pharmacol Ther 76 6 628 38 2004 Deng Blood 92 6 1981 1988 1998 Ji Hee Song Journal of Immunol Methods 286 187 201 2004 99 104940 000000 99 034950 000000
113. 10000 _ 5081 Selection Oigonucl eoti der DNA DO O GeneEdi tor Antibiotic Selection 0 D 000 ds DNA 0 2M NaOH 0 2 nMEDTAI 5 M pH4 6 10 000 Sel ecti on Oli eoti def 5 1000000 75 CO 500 0 0 0 0 37 CO 000 00000 100000 Synthesis 10x buffer T4 DNA Pol ynerase DNA ligase 37 90 GeneEdi tor Antibiotic Selection BMH 71 18 mt S UU DNA O DNAHHHHHHHHHHHJMO9HHHHHHH GeneEdi tor Anti bi oti Selection DNAD 0 5080 0000000000 Treo O hMpl 0 pEF MPL635 10 Mut Ser 508 5 CTGCTGCTGC TGAGGTCGCA GTTTCCTGCA CACTAC 3 00000 160 0 3 c MoIH L929H MPL635 1 90 Li pof ectam 0 Lipofectamne PLUSO Invitrogenl Dul becc
114. 3 13 M13 20FW 5 GTAAAACGACGGCCAGTG 3 14 1 5 CAGGAAACAGCTATGAC 3 15 28 02007 108559 Al 2007 9 27 PER i Yus C e r3 m r3 m T f Te r3 n 0 E mI r3 r3 16 H O FIL P b H H LU
115. 0 1 1nM 100000 100 10000000 1000000 lt 034 00000 0 CUM 00000 0 0 0 00 00 000 0000 000000000000000000000000000 Megacut qj Stem Cell chnol ogi es Cat 49720000000000000000000 152 02007 108559 Al 2007 9 27
116. 6 4 50 201 3 6 5 2 1801 2352010 2 DNP 3 8 8 8 o o 5 1 FDCP hMPL FDCP parent g 8 to 104 to 104 109 107 102 10 104 PE 102 103 104 10 107 102 10 104 10 102 10 5 E PE E 5 3 S 3 5 8 100 30 102 103 104 112 842 02007 108559 1 2007 9 27 e PEG UO 4 48 61 1 o o0 wu059 0St 00 24 7 10 Gt lt 14099 09 00 9 0001 0 01 1 100 0 000001 BE nM 4 9 6 5 2 61 0 01 100 0 0001 8 6 Ale en borra e ei Str 2 0 0 000001 PEG rHuMGDF _ 0 6 4 50 61 _____ 2 1039 09 40 0 nw Paol Feel 02007 108559 Al 2007 9 27 4 EcoRI EcoRY Sall 120 100 eU eue 80 PEG rHuMGDF P 60 7 10 61 n A 4 49 61 Ca 2 0 i 0 1 1 10 100 1000 10000 BE ng ml
117. 000 0000 DMEM 0 96 Ll 0 0000 d D 0 0 7 TPO BaF3 MolH Oita et al Blood 2005 15 105 2 562 6 0 U77TPO 00000 150 02007 108559 Al 2007 9 27
118. TPQ 0 0 000 10000000 h lt a b c d e f g h 133 02007 108559 1 2007 9 27 10000000000 1 1 I
119. A27_F 5 3 49 LV 3 202LR 5 AGAGAGAGAGCGTACGTTTAATCTCCAGTCGTGTCCCTTGGC 3 50 C C lee AE eee PIP PPIP An 20 43 02007 108559 Al 2007 9 27 6 4 50 HV 5 50 5 7Hsa1 5 AGAGAGAGAG GTCGACCACC ATGGAATTGG GACTGAGCTG GATTTT 3 51 HV 3 C15H3Nhe 5 AGAGAGAGAGGCTAGCTGAGGAGACGGTGACCGTGGT 3 52 6 5 2 10 HV 5 F24HSal 5
120. 4 496434411 DO 0 0 6 5 20000 x N 9STAT500000000000000001gc33440 Jak2 STAT5H BO ul 4 79 9 gt x 100000 100000000 3 1 0000000000000000000000000 1000000000 1409 15110000 0 atel et Rich Plasma PR PH 2 000000 0 0 25000 15mnHHHHHHHHHHHHHHHHHHHHH 3 PRPH 0 D 3 10 4 3 100000000 1001 L 5 30u SIGM O O O 5u LO 00 0 0000000000
121. DI DI DI DI DI DI DI DI 127 02007 108559 Al 2007 9 27 HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH 0000 0000000000000 1285 12E10 12D5H 10 00 Whole antibody 19910 000 D 10 100000000000000000000 000000000000000000000 0 0 0 0 t 10 HHHHHHHHHHHHHHHHHH CD34 TPO J
122. x 10000000000 000 hHHHHHHH H n n n n C JH ss x H Od 00000 000 2 UT7 TPO EC50 UT7 TPO Max 2 35 7 10 jgG1 gt 90 1 4 49 IgG1 gt 80 2 6 4 50 IgG1 gt 80 3 6 5 2 IgG1 gt 50 3 PEG rHuMGDF 0 001 0 01nM 100 1 10nM 0 1 1nM 0 000 n HH nn HH nnn HH nnm HH 10 20 30 40 50 33 WO2007 108559 Al 2007 9 27
123. 72 3 72 3 54 570000 Re a aE 1000000000000000000000000 EE Von Maps al D 00000000000000000000000 HHHHHHHHHHHHHHHHHHHHHHHHH 0000000000000000000000000 HHHHHHHHHHHHHHHHHHHHHHHHH IHHHHHHHHHHHHHHHHHHHHHHHH IHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHH IHHHHHHHHHHHHHHHHHHHHHHHH rr IHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHH Me lg 10 10 10 10 DAYS ELAPSED AFTER PRIMARY ADMINISTRATION 0000000000 00000000000000 HHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHH HHH HHHHHHHHHHHHHHHHHHHHHHHHH IHHHHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHH 02007 108559 Al 2007 9 27 2 10 20
124. DO HHH 000 LU 0 00 100000 uH 100000 00 100000 10 20 30 40 50 29 02007 108559 Al 2007 9 27 IU DO DO oa L3 0O n n n n n n n n 000 l 100 E 1 n n n 1000 00000000000 NH _MPSWALFMVTSCLLLAPQNLAQVSSQDVSLLASDSEPLKCFSRTFEDLTCFWDEEEAAPSGTYQLLYA YPREKPRACPLSSQSMPHFGTRYVCQFPDQEEVRLFFPLHLWVKNVFLNQTRTQRVLFVDSVGLPAPPSI IK AMGGSQPGELQISWEEPAPEISDFLRYELRYGPRDPKNSTGPTVIQLIATETCCPALQRPHSASALDQSPCA QPTMPWQDGPKQTSPSREASALTAEGGSCLISGLQPGNSYWLQLRSEPDGISLGGSWGSWSLPVTVDLPGDA VALGLQCFTLDLKNVTCQWQQQDHASSQGFFYHSRARCCPRDRYP IWENCEEEEKTNPGLQTPQFSRCHFKS RNDSIIHILVEVTTAPGTVHSYLGSPFWIHQAVRLPTPNLHWREISSGHLELEWQHPSSWAAQETCYQLRYT GEGHQDWKVLEPPLGARG
125. EcoRIH Xball 2k EFH HH Bsd 6 Neo pEGEP N1 LU 10000000000000000000000 Xbal 1000000000000 Alkaline Phosphat as e E coli C75 0 00000 p E EE B BB ee EE EE DE E BT EE E EH DNAD 0 0 DNI IU UU UU hip U U U DN 00000000060 T4 ligase 1000000000000 01080 DN O O DNA 00 0 MPL635 O pcw MPL635H Mol 5 AGAGAGAGAG GAATTCGCCA CCATGCCCTC CTGGGCCCTC 3 00000 120
126. AGAGAGAGAGGTCGACCACCATGGAGTTGGGACTGAGCTGGATTT 3 53 3 L66H3Nhe 5 AGAGAGAGAGGCTAGCTGAGGAGACGGTGACCGTGGTC 3 54 E C C ra ia HHHHHHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHH 20 30 1000000000 000000000000000000000000000000000000000 01000 40 1000000000 000000000 44 02007 108559 Al 2007 9 27 STKGPSVFPLAPCSRSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS SSLGTQTYTCNVNHKPSNTKVDKRVELKTPLGDTTHTCPRCPEPKSCDTPPPCPRCPEPKSCDTPPPCPRCP EPKSCDTPPPCPRCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFKWYVDGVEVHNA KTKLREEQYNSTFRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKTKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDI AVEWESNGQPENNYNTTPPMLDSDGSFFLYSKLTVDKSRWQQGNIFSCSVMHEA LHNRYTQKSLSLSPGK 55 1000000000 CTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGGAGCACCTCTGGGGGCACAGCGGCCC TGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCG GCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCT CCAGCAGTTTGGGCACCCAGACCTACACCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGA GAGTTGAGCTCAAAACCCCACTTGGTGACACAACT
127. 1 4 15 17 1 8 DNA DNA DNA DNA 1 4
128. 1 98 J 60 C 309 0 72 C30H 150 13ch1 linkH2H 0000 98 60 C 72 C300 0 0 B 0 1500000000 0 bNA PCR purification kit 00002 0 99 60 C30H 72 C 30 15000000000 linkHg linko 0000 150000 1000 DNA 10000 1961000000000 1000 10000 N5kG33119 0 0 li nkH GGG ACA CAG 570 13ch1 R GIC GIG O 0O O O 580 13ch1 GIG GIG GA 590 li nkH2 TGA TCA GAT 10000 600 5 1963331 0000000 19G3331 0 0 0 0 N5KG3H linkH CH3consRO 98 O 60 C300 72 30 10 150 0 N5KG1H CH3 cons IinkH2H 98 CH 60 0 72 C30H 00 0 0 15000000000 DN Aj O purification kit 2 0 0 0 00000000000 98 60 C30 72 30H 5 linkH IinkH2H 0000 15000 DNA Nnel Bami N5KG1H 19G1H 1000000000 N5KG3331 CH3consR GGTGTACACCTGTGGCTCTCGGGGCTGCCQ DO eir CH3cons GGGCAGCCCCGAGAGCCACAGG
129. 40 50 17m 1R 5 TGTGTGAGTT GTGTCACCAA GTGGGGTTTT CCTTGGT 3 72 17m 2F 5 ACCCCACTTG GTGACACAAC TCACACATGC CCACCATGCC CAGCACCTGA GTTCGAG 3 73 48 WO2007 108559 Al 2007 9 27 HH 00000 000 oe pa E ur dE RE Se RES SE T pr fica H mma m3 rm rA rA 10 20 30 40 50 49 02007 108559 1 2007 9 27 _ E NS nnnn
130. 68 C 1H 3500 100 72 C 7 N5KG4PE r 100 G3G4_P3 6364 DNA 10000 Q Gel Extraction Kit 100 DNAHHHHHHHHHHHHH 5 94 1001 94 C 100 55 10 68 C 1 500 000 100 00000 00000000 0 O0 O0 6364 P1 6364 94 C 50 68 C 2 10002000000 72 C 7 P1 FI 0304 RI 10000000000 P1 6364 PA PCR Q Aqui ck Gel Extraction KitH Zero Blunt TOPO PCR Cloning Kit 010 0000 0 0 00 0 per 4 Blunt 1000000000000000000 54100000000 54410000 10000000000 19633440 1963344111 10 _ 1_ 5 GCACCAAGGG CCCATCG 3 0 00 O 0 630 P2 5 GCTGGGCACC TTGGGCACG 3 00000 640 5 CCAAGGTGCC CAGCACCTGA GITCGAGGGG GGA 3 0 0 0 0O O 650 P4 5 AGAGAGGGAT 3 660
131. 75 ttggctattt gtacagcctg tatgccatgc agttggaata agagacaacg gccttgtatt gacgtctggg WO2007 108559 Al 2007 9 27 taaaaggtgt gcaggtccct actgggtccg gtggtagtat ccaagaactc actgtgcaaa gccaagggac ccagtgtgaa 60 gagactctcc 120 gcaagctcca 180 aggttatgcg 240 cctgtatctg 300 acctatatgg 360 cacggtcacc 420 429 Met Glu Leu Gly Leu Ser Trp Ile Phe Leu Leu Ala Ile Leu Lys Gly 1 Val Gln Cys Glu Val Gln Leu Pro Gly Arg Ser Leu Arg Leu 5 20 35 Asp Asp Tyr Ala Met His Trp 50 40 55 10 25 15 Val Glu Cys Gly Gly Gly Leu Val Gln 30 45 60 Ser Cys Ala Ala Ser Gly Phe Thr Phe Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met Asn 85 100 90 105 95 Ser Leu Arg Ala Glu Asp Thr Ala Leu 110 10 20 30 40 76 02007 108559 Al 2007 9 27 Cys Ala Lys Pro Trp Phe Gly Glu Trp Gly Asn Tyr Tyr 115 120 125 Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 130 135 140 lt 210 gt 37 lt 211 gt 387 lt 212 gt DNA lt 213 gt Homo sapiens lt 400 gt 37 atggaaaccc cagcgcagct tctcttcctc ctgctactct ggctcccaga taccaccgga gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt aagagccacc ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa c
132. E RI MELGLSWIFLLAILKGVQCEVQLVECGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSGISW NSGSIGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCAKP IWFGEWGNYYGMDVWGQGTTVTVSS 36 IDUU U 0000000000000000000000000000000000 00000000000 ATGGAAACCCCAGCGCAGCTTCTCTTCCTCCTGCTACTCTGGCTCCCAGATACCACCGGAGAAATTGTGTTG ACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAGGGCCAGTCAGAGT GTTAGCAGCAGCTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGATGCA TCCAGCAGGGCCACTGGCATCCCAGACAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACTCTCACCATC AGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTACTGTCAGCAGTATGGTAGCTCACCGATCACCTTCGGC CAAGGGACACGACTGGAGATTAAACGT 37 eB BE TE EE ET ED BE LE EET BEBE SE OH 0000000000000000000000000000000 METPAQLLFLLLLWLPDTTGEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYDA SSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPITFGQGTRLEIKR 38 GAL HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH a a 1000 100 1000000 HHHHHHHHHHHHHHHH HHH 000000000000
133. ST ATS 1 2 Akt Akt a Conor EJ 120 WO2007 108559 A1 2007 9 27 6B 6 5 2 Gl 6 5 2 G3344 PEG rHuMGDF 10ugimL lugmL 10OugmL lugmL 100ng mL 130k 200 70 280 2008 7 28 121 02007 108559 Al 2007 9 27 1 3 1 0000 2 ELKTPLGDTTHT 10 ESKTPLGDTTHT 11 CPRCP EPKSCDTPPPCPRCP x3 CPRCPEPKSCDTPPPCPRCP 5 CPR CPPCPH 100 0 UU OU b D a CD34 00 CFU 0000000 0 Q 10 000
134. ae a 30 i 50 02007 108559 Al 2007 9 27 10 10 20 30 40 l 50 11 02007 108559 Al 2007 9 27 20
135. linkH GGG GTC ACA AGT 57 13ch1 R GTC TTC GTG GCT CAC GTC CAC CAC GCA 58 13chl TGC GTG GTG GTG GAC AGC CAC GAA GAC 59 linkH2 TGA TCA GTA GAT ATC ACG GC 60 CH3consR GGTGTACACCTGTGGCTCTCGGGGCTGCCC 61 CH3cons GGGCAGCCCCGAGAGCCACAGGTGTACACC 62 EE li RC r NOE n C C C C C C C C C C m C 10 20 30 40 46 WO2007 108559 1 2007 9 27 G3G4_P
136. lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 47 agagagagag atctctcacc atggacatga gggtccccgc tcagc lt 210 gt 48 lt 211 gt 40 212 DNA lt 213 gt Artificial Sequence WO2007 108559 Al 2007 9 27 10 41 20 42 30 45 40 80 lt 220 223 gt Description of Artificial Sequence primer lt 400 gt 48 agagagagag cgtacgtttg atttccacct tggtcccttg lt 210 gt 49 lt 211 gt 47 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 49 agagagagag atctctcacc atggaaaccc cagcgcagct tctcttc 210 50 lt 211 gt 42 lt 212 gt DNA lt 213 gt Artificial Sequence 220 223 Description of Artificial Sequence primer lt 400 gt 50 agagagagag cgtacgttta atctccagtc gtgtcccttg gc 210 51 lt 211 gt 46 212 DNA X213 Artificial Sequence 220 lt 223 gt Description of Artificial Sequence primer 400 51 agagagagag gtcgaccacc atggaattgg gactgagctg gatttt JP WO2007 108559 1 2007 9 27 40 10 47 20 30 42 40 46 81 lt 210 gt 52 lt 211 gt 37 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 52 agagagagag gctagctgag gagacggtga ccgtggt lt 210 gt 53 lt 211 gt 45 lt 212 gt DNA lt 213
137. lt 220 gt lt 223 gt Description of Artificial Sequence primer WO2007 108559 1 43 18 17 2007 9 27 10 20 30 40 64 lt 400 gt 16 ctgctgctgc tgaggtcgca gtttcctgca cactac lt 210 gt 17 lt 211 gt 491 lt 212 gt PRT lt 213 gt Homo sapiens lt 400 gt 17 Met Pro Ser Trp Ala Leu Phe Met Val Thr Ser Cys 1 5 10 Pro Gln Asn Leu Ala Gln Val Ser Ser Gln Asp Val 20 25 Ser Asp Ser Glu Pro Leu Lys Cys Phe Ser Arg Thr 35 40 Thr Cys Phe Trp Asp Glu Glu Glu Ala Ala Pro Ser 50 55 60 Leu Leu Tyr Ala Tyr Pro Arg Glu Lys Pro Arg Ala 65 70 75 Ser Gln Ser Met Pro His Phe Gly Thr Arg Tyr Val 85 90 Asp Gln Glu Glu Val Arg Leu Phe Phe Pro Leu His 100 105 Asn Val Phe Leu Asn Gln Thr Arg Thr Gln Arg Val 115 120 Ser Val Gly Leu Pro Ala Pro Pro Ser Ile Ile Lys 130 135 140 Ser Gln Pro Gly Glu Leu Gln Ser Trp Glu Glu 145 150 155 Ile Ser Asp Phe Leu Arg Tyr Glu Leu Arg Tyr Gly 165 170 WO2007 108559 Al 2007 9 27 Leu Leu Leu Ala 15 Ser Leu Leu Ala 30 Phe Glu Asp Leu 45 Gly Thr Tyr Gin Cys Pro Leu Ser 80 Cys Gln Phe Pro 95 Leu Trp Val Lys 110 Leu Phe Val Asp 125 Ala Met Gly Gly Pro Ala Pro Glu 160 Pro Arg Asp Pro 175 10 20 30 40 65 WO2007 108559 Al 2007 9 27 Lys Asn Ser Thr Gly Pro Thr Val Gln Leu Ile Ala Thr Glu Thr 180 185 190 Cys Cys Pro Ala Leu Gln Arg Pro His Ser Ala Ser Ala Leu Asp Gln 195 200 205
138. 003 1oux 1954 ed grz mas EAA FE ag 6903 FR weg AL EAS 12828955 haog Tajo HEI IEA 19ASN i 12458 pud 1169 1984 14828955 EL HEU AILES 1auN 12053 OIE CA vndn1 uI kd d 824 AH XXX nes 2 HH ddd AT TI Misa IAMSB ibe 1 02007 108559 Al 2007 9 27 115 peri M iE 3 c c3 m pn 3 m 2200222 5555555 3335355353 5555555 SESSSEE 55553555 SSSSSee FERRARA EEEEEEE EEGPEGR SARRERA 2222202 2222222 EEEEEEE 5555555 EEEEEEE 2222233 22258 TIU EEESEEET 5255255 55855585 94 2222222 55595595 2222222 2225225 996199196 LLL 802 5588555 555555 3838353 S555555 BEEBE TB SSSSSSS 2222225 samasaa 2222222 2222222 IHueg 2022292 SESESBS 2900990523 2222202 3445448 TEETTTT 2266686 8555555 18 4 H N H 1 V 3 H H A S 2 S 4 A EEEE
139. 100000000 98 C 109 50 C 300 72C 57000 98 C 100 55 C 300 0 0 0 0 O 7290 10 5 98 10 68 C 1 10 000000 0 25000000000 72 7 Pyrobest DNA Pol 1000 000 N5KG3344h1H 100000000 17 22 70 cse PAR H Hl 0 D PCR DNAHHHHHHHHHHHHH ck Gel Extraction 1000000000 DAD 00000 DNA O 1000000000000000 98 00 10 0 98 C 1 d dg ee c 1gn 10500 98 C 10 55 30 72 C 10 10 500 100000000 1000000000000 6434 P5 6364 O O 94 C 30 68 19 0 0000003000 72 739 00000000 10000 PCR 1000000000000 Q Aqui Gel Extraction Kit 10000000 pCR 4 nt TOPOHHHHHHHHHHHHHHHHHHH DNAIH DNAHHHHHHHHHHHHHH 1gG4344uh HHHHHHH 17 180 5 AGGTGCTGGG CACCGTGGGC ATGTGTGAGT 3 00000 700 17 2 5 CACACATGCC CACGGTGCCC TTC 3 D H D 710 100000 10 gG4344uhnh UD gG4344uhnf NSKG4PEH 34_P5_F 17m 1 98 100 50 30 729 1 159 0000000 98 C 109 55 0 0 0 72 1 500000 98 100 0 0 0 0000000 2500 0 72 C 7 PCR Pyrobest DNA Pol
140. 19290 0 0 00 0 O 929 FACS Vantage Be cton 0 0 000 000000 0 0 9 Blasticidin 10 BSO 0 DMEM 10000000 4 0000 lt 0 330000000 MPL635 0 FMBAH c MolH 0 FM 0 00 5bM g m Blasticidin 10 BS Roswell P ark Memorial Institute 550000000 FMBAU MPL635 Ser 3 FMBAI hMol FMGA Ser 5u g m Blasticidin 10 BSO RP M 1 2 c MID c MIQO 000000 LUTO DNAH pEAK8 EdgeBi oSystems Promega Hek293 10000000 000000000 soluble MI x sMl x 10 NH MPSWALF M TSCLLLAPQNLAQVSSQDVSL LASDSEPLKCF SRTF EDLT CF WDEEEAAPSGTYQLLYAYPREKPRA CPLSSQSMPHF GTRYVCQF PDQEEVRLFFPLHLVWKNVFLNOTRT QRVLFVDSVGLPAPPSI KAMGGSQPGELQI SWEE PAPEI SDFLRYELRYGPRDPKNSTGPTVI QLI ATETCCPALQRPHSASAL DQSPCAQPT MPWQDGP KOTSPSREASALTA EGGSCLI SGLQPGNSYW QL RSEPDGI SLGGSWGSVSLPVTVDLPGDAVALGL QCF TL DLKNVT CQWQQQDHASS FY HSRARCCPRDRY PI MENCEEEEKTNPGLQTPQF SRCHF
141. 41 bone morphogenic proteing 10 0 20 0 30 0 00 40 5 6 7 8 9 100 0 0000 110 12 00000 13 140 00000 15 I a 10 1100 2a JUD 20 J B 0 0 11000 100000000 4010000000005 6 1007 808 000000000 800 8 1000009 00 0 10 10000 2 related protein 0 1000000000 000000000 00000001 3 00 4 2 10000000000 ABH HH BI BB 0 1100009000000 0 0 TNFQ TNF1H TNF2 HHHHHHoHHHHH 10 B 1H B 1 20 10 20 000000 8 3 5 latent 100000 1000000 1000000000 10000 10000 EN 10000000000100000000000000000 10000000 146 02007 108559 Al 2007 9 27
142. C 3 00000 220 1000 7 100000000000000000000000 2400000 00000000000000000000000000000000 lt 7 100 0 0 000 0 000000000090000000000000 ATGGAGTT GGGACT GAGCT GGATTTT CCTTT TAAAAGGT GT CCAGT GT GAAGT GCAGCT GGT GGAGT CTGG GGGAGGCTT GGT ACAGCCT GGCAGGT CCCT GAGACT CT CCT GT GCAGCCT CT GGATT CACCT TT GATGATT AT GCCAT GC ACT GGGT CCGGCAAGCT CCAGGGAAGGGCCT GGAGT GGGT CT CAGGTAT GGAATAGT GGTAGCATAGGCT AT GCG GT GAAGGGCCGATT CT CCAGAGACAACGCCAAGAACT CCCT GTAT CT GCAAAT GAACAGT CT GAGAGC TGAGGACACGGCCTT GTATTACT GT GCAAAAAAT CTAT GGT TCGGGGAGTTCCGTTACTGGTACTTCGATCTCTGGGGCC GTGGCACCCTGGTCACTGTCTCCTCA DDD 230 lt 7 10 JU UU DU 0 0000000000000000 HHHHHHHHHHHHHHHHHHHHHHHHHHHILH MELGLSW FLLAI LKGVQCEVQLVESGGGLVQPGRSLRL SCAASGF TF DDYAMHVWRQAP GKGL EW SGI SWNSGSI GYA DSVKGRFTI SRDNAKNSLY L QVNSLRAEDTALYY CAKNL GEFRYW FD LWGRGTLVTV SS 0 0 240 lt 7 1000 1000 ATGGACATGAGGGTCCCCGCTCAGCTCCTGGGGCTTCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCATCCAGTTGAC CCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGTG CTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTC CCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAAC
143. Cat 93520 l phospho STAT50 Cell Sig nal i O Cat 9351 JAK2 Upstate O Cat 206 255 J 2 Upst ate Cat 407 606 Erk1 20 Cell Signaling D Cat 9272 phospho 1 20 Cell Signaling Cat 92711 D Akt Cell Signaling Cat 91020 pho spho Cell Signaling O Cat 910150 100000 1000000000000000000000000 1 UT7 TPO IMDM 2 0000000 1x 106cells ntH 6 wellH 2nL wellH 000 0 3 HHHHHHHHH PEGrHuMGDFH 00000 166 02007 108559 1 2007 9 27 4 5 HHHHHHHHHHHHHHHHHHHHH PhosphoSafe Extraction Reagen Cat 71296 100000 6 0050000000000000000000000000000000000 YQ 7 1064344 1000000000000000000
144. Ser Pro Cys Ala Gln Pro Thr Met Pro Trp Gln Asp Gly Pro Lys Gln 210 215 Thr Ser Pro Ser Arg Glu Ala 225 230 Cys Leu Ile Ser Gly Leu Gln 245 Arg Ser Glu Pro Asp Gly Ile 260 Ser Leu Pro Val Thr Val Asp 275 Leu Gln Cys Phe Thr Leu Asp 290 295 Gin Gln Asp His Ala Ser Ser 305 310 Arg Cys Cys Pro Arg Asp Arg 325 Glu Glu Lys Thr Asn Pro Gly 340 His Phe Lys Ser Arg Asn Asp 355 Thr Thr Ala Pro Gly Thr Val 370 375 Ile His Gln Ala Val Arg Leu 385 390 220 Ser Ala Leu Thr Ala Glu Gly 235 Pro Gly Asn Ser Tyr Trp Leu 250 Ser Leu Gly Gly Ser Trp Gly 265 270 Leu Pro Gly Asp Ala Val Ala 280 285 Leu Lys Asn Val Thr Cys Gln 300 Gln Gly Phe Phe Tyr His Ser 315 Tyr Pro Ile Trp Glu Asn Cys 330 Leu Gln Thr Pro Gln Phe Ser 345 350 Ser Ile Ile His Ile Leu Val 360 365 His Ser Tyr Leu Gly Ser Pro 380 Pro Thr Pro Asn Leu His Trp 395 Gly Ser 240 Gln Leu 255 Ser Trp Leu Gly Trp Gln Arg Ala 320 Glu Glu 335 Arg Cys Glu Val Phe Trp Arg Glu 400 10 20 30 40 66 02007 108559 1 2007 9 27 Ser Ser Gly His Leu Glu Leu Glu Trp Gln His Pro Ser Ser Trp 405 410 415 Ala Ala Gln Glu Thr Cys Tyr Gln Leu Arg Tyr Thr Gly Glu Gly His 420 425 430 Gln Asp Trp Lys Val Leu Glu Pro Pro Leu Gly Ala Arg Gly Gly Thr 435 440 445 Leu Glu Leu Arg Pro Arg Ser Arg Tyr Arg Leu Gln Leu Arg Ala Arg 450 455 460 Leu Asn Gly Pro Thr Tyr Gln Gly Pro Trp Ser S
145. WO2007 108559 Al 2007 9 27 Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys 340 lt 210 gt 75 lt 211 gt 377 lt 212 gt PRT lt 213 gt Artificial Sequence lt 220 gt 345 lt 223 gt Description of Artificial Sequence 63344 lt 400 gt 75 Ala Ser Thr Lys Gly Pro Ser 1 5 Ser Thr Ser Gly Gly Thr Ala 20 Phe Pro Glu Pro Val Thr Val 35 Gly Val His Thr Phe Pro Ala 50 55 Leu Ser Ser Val Val Thr Val 65 70 Tyr Thr Cys Asn Val Asn His 85 Arg Val Glu Leu Lys Thr Pro 100 Arg Cys Pro Glu Pro Lys Ser 115 Cys Pro Glu Pro Lys Ser Cys 130 135 Val Phe Ala Leu 25 Ser Trp 40 Val Leu Pro Ser Lys Pro Leu Gly 105 Cys Asp 120 Asp Thr Pro Leu Ala Pro Cys Ser Arg 10 15 Gly Cys Leu Val Lys Asp Tyr 30 Asn Ser Gly Ala Leu Thr Ser 45 Gln Ser Ser Gly Leu Tyr Ser 60 Ser Ser Leu Gly Thr Gln Thr 75 80 Ser Asn Thr Lys Val Asp Lys 90 95 Asp Thr Thr His Thr Cys Pro 110 Thr Pro Pro Pro Cys Pro Arg 125 Pro Pro Pro Cys Pro Arg Cys 140 Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg Cys Pro 145 150 155 160 10 20 30 40 Ala Pro Glu Phe Glu Gly Gly Pro Ser 165 Pro Lys Asp Thr Leu Met Ile Ser Arg 180 185 Val Val Asp Val Ser Gln Glu Asp Pro 195 200 Val Asp Gly Val Glu Val His Asn Ala 210 215 Gln Phe Asn Ser Thr Tyr Arg Val Val 225 230 Gln Asp Trp Leu Asn Gly Lys Glu Tyr 245 Gly Leu Pro Ser Ser Ile Glu Lys Thr 260 265 Pro Arg Glu
146. s Modified Eagle DEM OO 0000000000 1 5x10 cells well H 6 wl plate OOOO 19290 0000000000000000000 01000001 148 02007 108559 Al 2007 9 27 0 0 0 U FBS 0 0 0 0 00 00 0 0 0 0 0 0 0 00 00 10 gm Blast Invitrogenl HHHHHHHHHHHHHHHHHHHHHHHHc MIHH Fl uorescence Activated Cell Sorting FACS 0 c MolH c
147. 13 WD2007 108559 1 2007 9 27 901191010100 1000000000 CR MET NH E EM CRM 10 20 30 40 50 02007 108559 Al 2007 9 27 14 10 20 30 40 50 1000
148. EC50 1nM IU LU 173 02007 108559 Al 2007 9 27 7 O00 1 0 8
149. Ud I 10 000 1 000 PBS 111 10 000 7 1064344 1001 g head week I V 1 000 7 10G4344uhni 1001 9 head week V 10 000 TPO PEG r HuMGDF O 5p 9 week VI 1 000 TPO PEG r HuMGDF 5 g head week HHHHHHHHHHHHHHHHHHHHHHHHHHH O l O 000 709 0000 2 5 10000 00 000000 FACS 0 00000000 CD41 Dako R7058 O jp sm CD41 BD Pharm ngen 0 5538487 000000000 00000 000 CI UU M2473 CD45 BD Pharmingen 9 553080 U 0000 000 1000000000 0000 HHHHHHHHHHH 0 O O TPQ 50ng SCF 100ng ni 0 37 C 5x00 0 000 0 CD41 0 Methocult systenf Stem Technol ogi es EPO 41 U m SCF 100ng IL 3 20ng nt 0 GM CSF 10ng ni 37 5 590 au uugullilululiuilddiiililiillibilii liuiuiuilWi ilililiu6tL i lilib iu l do
150. attttgcaac ccaagctgga sapiens 135 tcagctcctg ccagtctcca aagtcagggc gctcctgatc tggatctggg ttattactgt gatcaaacgt Eggcttctgc tcctccctgt attagcagta tatgatgcct acagatttca caacagttta 140 tgctctggct ctgcatctgt ctttagcctg ccagtttgga ctctcaccat atagttaccc cccaggtgcc 60 aggagacaga 120 gtatcagcag 180 aagtggggtc 240 cagcagcctg 300 gtacactttt 360 390 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 20 30 40 72 02007 108559 Al 2007 9 27 Leu Pro Gly Ala Arg Cys Ala Gln Leu Thr Gln Ser Pro Ser Ser 20 25 30 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45 Gln Gly Ile Ser Ser Thr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60 Ala Pro Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val 65 70 75 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 85 90 95 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 100 105 110 Phe Asn Ser Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile 115 Lys Arg 130 lt 210 gt 31 lt 211 gt 423 lt 212 gt DNA lt 213 gt Homo lt 400 gt 31 atggaattgg gtgcagctgg tgtgcaacct gactctgtga caaatgaaca ttcggggagt tca sapiens gactgagctg tggagtctgg ctggattcac tggagtgggt agggccgatt gtctgagagc tccactacgg 120 gattttcctt gggaggcttg ctttgataat ctcaggtatt caccatctcc tgaggacacg tctggacgtc ttggctattt gtacagcctg
151. 0000 2160 2260 226 231 gG3 100000000000000000000000000 G3uh upper hinged OO 00 0 0 0 I gGx3xxuh 00000 0 G3uh 12175 22 G3uh 1gG4PEU 00 00000000000 0 O O Gunni upper hinge mt ation 19G3xxuhr 0 1000000000000 19 1964344 1gG4344h1 gG4344uh 158 02007 108559 Al 2007 9 27 gG4344uhnf MIHHHHHHHHHHHHHHHHHHHHHHH 1000000000000 100000 1941 gG4PE 1963311 1903331 1963344 196334411 1904344 gG4344h1 196434 4uh gG4344uhm 1
152. 021 000014 OFI 00070 ver 10 9 9 710G4344uhm_3u 9 710G4344uhm 10u 600 500 02 NGAN 4 t 90 0 gt d i 00041 ERE P G g 8 E a 5 E WAS Sao ei 14 IN 13 19 146 amp ab m 2 1 Odl OM 143 i i i 60 0 gt 000 01 o o N A 0 118 02007 108559 1 2007 9 27 00000 11 E12 14 7 10G4344U 250 Ly V104L TH 4 1 A43V 1 200 lt LV 01000 HNP 191 1 150 a 5 8 0 8 J 100 Fe L104V c 06 A43V 50 64000 04 lt 710G4344uhm 0 0 0 01 0 1 1 02 ug mL 0 0 0 00001 0 001 0 1 10 1000 nM 119 02007 108559 A1 2007 9 27 00000 6 A 7 10 4 49 PEG G4344uhm G4344uhm rHuMGDF lug mL lugmL 100ngmL 0 5 30 0 5 30 0 5 30 JAK2 JAK2 STATS STATS Erkl 2 Erk1 2 Akt Akt 00000 JAK2 JAK2 5
153. 1 5 17 1 8 DNA PCT 1 3 2 1 13 14 15 17 18 19 1 4 15 17 1 8 1 4 15 17 1 8 4 4 8 8 DNA G 1
154. 10000 STKGPSVF PLAPCSRSTSGGTAAL GCL VKDY F PEPVT VSVNSGAL TSGVHTF PAVL QSSGLYSLSSVVTVPSSSLGT QTY 161 02007 108559 1 2007 9 27 TCNVNHKPSNTKVDKRVELKTPLGDTTHTCPRCPEPKSCDTPPPCPRCPEPKSCDTPPPCPRCPEPKSCDTPPPCPRCPA PELLGGPSVFLFPPKPKDTLM SRTPEVTCVVVDVSHEDPEVQF KWYVDGVEVHNAKTKLREEQYNSTFRVVSVLTVLHO DWLNGKEYKCKVSNKALPAPI SKTKGQPREPQVYTLPPSREEMIKNQVSLTCLVKGFYPSDI AVEWESNGQPENNY NTTPPM_DSDGSFFLYSKLTVDKSRWOQGNI FSCSVMHEALHNRYTOKSLSLSPGK 0 0 0 550 19630 0000000 CTAGCACCAAGGGCCCAT CGGT CTT CCCCCT GGCGCCCT CCAGGAGCACCT CT GGGGGCACAGCGGCCCT GGGCT GC CTGGTCAAGGACTACTTCCCCGAACCGGT GACGGT GT CGTGGAACT CAGGCGCCCT GACCAGCGGCGT GCACACCTTCCC GGCT GT CCTACAGT CCTCAGGACT CT ACT CCCT CAGCAGCGT GGT GACCGT GCCCT CCAGCAGTTT GGGCACCCAGACCT ACACCTGCAACGT GAAT CACAAGCCCAGCAACACCAAGGT GGACAAGAGAGTT GAGCT CAAAACCCCACTT GGTGACACA ACTCACACAT GCCCACGGT GCCCAGAGCCCAAAT CTT GT GACACACCT CCCCCGT GCCCACGGT GCCCAGAGCCCAAATC TT GT GACACACCTCCCCCAT GCCCACGGT GCCCAGAGCCCAAATCTTGT GACACACCT CCCCCGT GCCCAAGGTGCCCAG CACCTGAACT CCT GGGAGGACCGT CAGTCTT CCT CTT CCCCCCAAAACCCAAGGATACCCTT AT GATT T CCCGGACCCCT GAGGTCACGT GCGT GGT GGT GGACGT GAGCCACGAAGACCCCGAGGT CCAGTT CAAGT GGT ACGT GGACGGCGT GGAGGT GCAT AAT GCCAAGACAAAGCT GCGGGAGGAGCAGT ACAACAGCACGTT CCGT GT GGT CAGCGTCCT CACCGT CCT GCACC AGGACTGGCT GAACGGCAAGGAGTACAAGT GCAAGGT CT CCAACAAAGCCCT CCCAGCCCCCATCGAGAAAACCATCTCC AAAACCAAAGGACAGCCCCGAGAACCACAGGT GT ACAC
155. 10000 4000 0 10m 0 00 H OOO50 00000000 37 2000000000000 0 5000000000000 00000 8 0 000000000 00000000 000000000 0000000000000000000 HGPRI 0000000000000 138 02007 108559 1 2007 9 27 0 210000000000000000000000000000000000 10000010000000000000000000000000 JPPH HDH 100100000000000000000000000000001 um DOU000
156. 19 JP 21 8 6 2009 8 6 51 Int Cl C12N 15 09 2006 01 CO7K 16 28 2006 01 CO7K 16 46 2006 01 A61K 39 39 2006 01 A61P 70 2006 01 2008 506356 P2008 506356 21 PCT JP2007 056536 22 19 3 20 2007 3 20 11 4294082 P4294082 45 21 7 8 2009 7 8 31 2006 81322 P2006 81322 32 33 31 JP 33 JP J WO2007 108559 Al 2007 9 27 AI 11 WO2007 108559 43 19 9 27 2007 9 27 FI C12N 15 00 ZNAA 48024 CO7K 16 28 48064 CO7K 16 46 40085 A61K 39 395 4HO45 6 7 00 18 3 23 2006 3 23 2006 299554 P2006 299554 32 18 11 H2H 2006 11 2 A 184 71 000001029 1 6 1 74 100091096 74 100096183 AK 74 100118773
157. 3 DI 3 DI DI DI DI DI 129 WO2007 108559 Al 2007 9 27 coli UichH Blood 86 971 976 1995 0 00000000 10000000000000000000000000000000000000 PEG HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHIH PEG NH SPAPPACDLRVLSKLLRDSHVLHSRLSQCPEVHPLPTPVLLPAVDFSLGEWKTOMEETKAQDI LGAVTLLLEG VMAARGQLCPT CLSSLLGQLSGQVRLLLGAL QSLLGT QLPPQGRTTAHKDPNAI FLSFOHILLRGKVRFLM_LVGGSTLCVR RAPPTTAVPS COOH 1 5 1 UT7 TPO HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH 10 000ng ni 1000000 000ng 0 0 0000000000000000 100ng 00000 1000000 uu 0
158. 30 40 62 WO2007 108559 Al 2007 9 27 lt 210 gt 10 lt 211 gt 12 212 gt PRT lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence mutated hinge Domain UH1G3uh lt 400 gt 10 Glu Leu Lys Thr Pro Leu Gly Asp Thr Thr His Thr 1 5 10 lt 210 gt 11 lt 211 gt 12 lt 212 gt 213 gt Artificial Sequence lt 220 gt 223 gt Description of Artificial Sequence mutated hinge Domain UH2G3uhm lt 400 gt 11 Glu Ser Lys Thr Pro Leu Gly Asp Thr Thr His Thr 1 5 10 lt 210 gt 12 lt 211 gt 42 lt 212 gt DNA 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 12 agagagagag gaattcgcca ccatgccctc ctgggccctc tt 42 lt 210 gt 13 lt 211 gt 43 10 20 30 40 63 lt 212 gt DNA 213 gt Artificial Sequence lt 220 gt 223 gt Description of Artificial Sequence primer lt 400 gt 13 agagagagag cggccgctca aggctgctgc caatagctta gtg lt 210 gt 14 lt 211 gt 18 212 DNA 213 gt Artificial Sequence 220 lt 223 gt Description of Artificial Sequence primer 400 14 gtaaaacgac ggccagtg lt 210 gt 15 lt 211 gt 17 212 DNA lt 213 gt Artificial Sequence 220 lt 223 gt Description of Artificial Sequence primer lt 400 gt 15 caggaaacag ctatgac lt 210 gt 16 lt 211 gt 36 lt 212 gt DNA lt 213 gt Artificial Sequence
159. 3m Nu 1 100 1000 100070 1000070 00L 1070 r 0 a T 120 0 T 50 e CS v Tues 30 gnbbepo 01 2 1 oe EMI 8 A E 0 3 9 H N S 1482 9881156 HIH 212 122 221 944 991 ILL 191 3211994 LOL 989 BOO SOY 2UH 211 298 969 la993 d S S 7 9 L A 3 9 V A H M 3 A 982 000 031 JHO 922 210 618 OOL LLI 996 993 014 OUD 111 919 111 212 913 218 222 601 298 612 399 082 182 298 219 BB9 391 220 281 219 383 261 389 19 V A S 11 14 5 98 18 A 1 8 d 3 S 0 194981 102 LLL 29 201 389 991 921 OUO 991 ALU 994 921 012 211 994 919 219 212 L L 839 298 913 OLI 328 20H JOH 313 26H JHL 308 JJH 208 286 983 OUO 896 5 3 0 8 N 9 5 b Y N 3 A D A 3 3 429 982 HIL 915 319 994 911 622 29H 119 969 909 911 HLI 383 211 238 219 189 111 999 212 219 199 949 221 IU 199 DIL HEI 319 229 190 915 OHH 901 982 519 200 985 COS A d N N 1 13 AS v L 9 S 3 3 qe 121 909 9BH 911 ULL 9H9 289 938 HH 952 122 LLL 219 212 HOY 222 161 211 388 184 913 912 291 519 119 LIL 338 109 899 LOL 911 069 949 LUO Zeb 5 d d 31 3 A S d YYA 1 3 3 A A 1 9 zl 199 399 983 199 192 291 599 LLL 981 21
160. 75 80 Tyr Thr Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val Glu Leu Lys Thr Pro Leu Gly Asp Thr Thr His Thr Cys Pro 100 105 110 10 57 20 30 40 90 02007 108559 Al 2007 9 27 Arg Cys Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg 115 120 Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val 130 135 140 Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 145 150 155 Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu 165 170 Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 180 185 Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser 195 200 Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys 210 215 220 Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile 225 230 235 Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 245 250 Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 260 265 Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 275 280 Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser 290 295 300 125 Phe Leu Phe Pro Pro Glu Val Val Gln Phe 175 Thr Lys Pro 190 Val Leu Thr 205 Cys Lys Val Ser Lys Ala Pro Ser Gln 255 Val Lys Gly 270 Gly Gin Pro 285 Asp Gly Ser Thr 160 Asn Arg Val Ser Lys 240 Glu Phe Glu Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly 305 310 315 320 Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 325 330 335 10 20 30 40 91
161. 94 15000000 0 98 C 100 68 C 3U 0000 30 72 C 7 PCR HH li D EP Q Aqui Gel Extraction 0 DNA O O pCR4 Bl unt 0 DNAD 00000000 ML3 20FVW 0 ML3RV 000000000 9544000000000 147 WO2007 108559 Al 2007 9 27 hMol 1 GenBank ACCESSION M901020 1000 DNAH hMIHHH DNA
162. 991 212 919 919 112 909 912 212 989 199 299 218 008 JLH 060 019 DIMILILILBOH 1169 60 E 7 A PEG rHuMGDF 7 10G4PE lt B PEG rHuMGDF 7 10G3344h1 1000 Ca i leuo t 1 1 1 1 L 10 0 SOE 7 88 9 10 11 12 13 1 2 3 4 5 6 LO 1 le m 4 lo N Ap 1 A don o gt N 23 c 9225958 ET5 5 55 ari har oon 5 0255 2299588888888 999999 9 sf 00 dE E k k db dE t 02007 108559 Al 2007 9 27 117 10 0 gt 4 000 1 e 0001 041 0001 201 2 0001 IGAN 1 GURNA 45 e Y S SM 0 he u LI 5 J o gu 9 04 009 oe 000 01 6 1n 201 WITH lt 00001 041 00001 2 2
163. ASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPYTFGQGTKLEIKR 30 E tN 0 00000000000000000000000000000000000000 U0 0 0U000000000000000000000000000000000 HHHHHHHHHHHH 10 20 30 40 37 02007 108559 Al 2007 9 27 ATGGAATTGGGACTGAGCTGGATTTTCCTTTTGGCTATTTTAAAAGGTGTCCAGTGTGAAGTGCAGCTGGTG GAGTCTGGGGGAGGCTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCAACCTCTGGATTCACCTTT GATAATTATGCCATGTACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAGTGGGTCTCAGGTATTAGTTGG AATAGTGGTGACATAGGCTATGCGGACTCTGTGAAGGGCCGATTCACCA TCTCCAGAGACAACGCCAAGAAC TCCCTGTATCTGCAAATGAACAGTCTGAGAGCTGAGGACACGGCCTTGTATTACTGTGCAAGGGATGCGGGG TTCGGGGAGTTCCACTACGGTCTGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA 10 31 U0 0 0U00000000000000000000000000 IO0U0O00000000000000000000000000050 MELGLSWIFLLATLKGVQCEVQLVESGGGLVQPGRSLRLSCATSGFTFDNYAMYWVRQAP GKGLEWVSGISWNSGDIGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCARDAG FGEFHYGLDVWGQGTTVTVSS 32 I0 0 00000000000000 0000000000000 ATGGACATGAGGGTCCCCGCTCAGCTCCTGGGGCTTCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCATC 1000000000000 20 1000000 CAGTTGACCCAGTCTCCATCCTCCCTGTCTGCAT
164. Ile 35 40 45 10 20 30 40 110 02007 108559 1 2007 9 27 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Phe Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 210 89 lt 211 gt 11 lt 212 gt PRT lt 213 gt Homo sapiens lt 220 gt 222 9 9 lt 223 gt Xaa is Ala or Thr lt 400 gt 89 Arg Ala Ser Gln Gly Ile Ser Ser Xaa Leu Ala 1 5 10 lt 210 gt 90 lt 211 gt 7 lt 212 gt PRT lt 213 gt Homo sapiens lt 400 gt 90 Asp Ala Ser Ser Leu Glu Ser 1 5 10 20 30 40 111 lt 210 gt 91 419 9 lt 212 gt lt 213 gt Homo sapiens lt 220 gt lt 222 gt 8 8 lt 223 gt is Leu Tyr or Trp lt 400 gt 91 Gln Gin Phe Asn Ser Tyr Pro Xaa Thr 1 5 lt 210 gt 92 211 gt 12 lt 212 gt lt 213 gt Homo sapiens lt 400 gt 92 Arg Ala Ser Gln Ser Val Ser Ser Ser Tyr Leu Ala 1 5 10 lt 210 gt 93 lt 211 gt 7 lt 212 gt lt 213 gt Homo sapiens lt 400 gt 93 Asp Ala Ser Ser Arg Ala Thr 1 5 lt 210 gt 94 lt 211 gt 9 lt 212 gt PRT WO2007 108559 Al 2007 9 27 10 20 30 40 lt 213 gt Homo sapiens lt 400 gt 94 Gln Gln Tyr Gly Ser Ser Pro Ile Thr 1 rode 8 4 49 1201
165. KSRNDSI LVEVTTAPGTVHSYLGSPFW HOAVRLPTPNL HMREI SSGHLELEWOHPSSWAAQET CY QL TGEGHQDVKVL EPP L GARGGT L ELRP RSRY RLQLRARL NGPT Y QGP VES VWSDPTRVETATETAW 17 0000 000 KM TM KM K Ul 1900000 14 5020 0 D igx OOOO D 5 KM 19000 K Hood 19 000 gk lg K 140 sc20 0 000000000 0 et al Proc Natl Acad Sci USA 2000 Vol 97 7220 gk KCo5H
166. LA LG LK LR LS LT LU LY M MG K MN W NG NQ NZ PG PH PL PT RS RJ SC SD SE SG SK SL SM SV SY TM TN TT TZ UA UG U5 G W ZA ZM ZW 72 IHHHHHHHHHHH HHHHHHHHHHHH 72 HHHHHHHHHHHH 00000000000 000000 72 1000000 00000000000 000000 48024 AAO1 7 2 4B064 07 CA9 G24 DOL 40085 8812 0102 O82 062 44045 AO BA57 DN6 2
167. PEG rHuMGDF 50 0000000 70 10000009 0 50 0 H 100nM 10nM 1nM HHHHHHHHHHH 2 9 10 000 ng ni b UT7 TPO HHHHHHHHHHHHHHHHHHHHHHH PEG r Hu MGDF 50 5000 EC50 100nM LU 000 11 6 0000000000 000000000000 0 CD34
168. Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 105 lt 210 gt 6 lt 211 gt 122 lt 212 gt PRT lt 213 gt Homo sapiens lt 400 gt 6 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 Ser Leu Arg Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe Asp Asn Tyr 20 25 02007 108559 Al 2007 9 27 30 95 15 Ala Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Gly Asp Ile Gly Tyr Ala Asp Ser Val 50 55 60 Lys 1 Arg Phe Arg Asp Asn Lys Leu 65 70 15 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Arg Asp Ala Gly Phe Gly Glu Phe His Tyr Gly Leu Asp Val Trp 100 105 Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 lt 210 gt 7 lt 211 gt 108 lt 212 gt PRT 110 10 20 30 40 lt 213 gt Homo sapiens 400 gt 7 Ala 6ln Leu Thr 1 Ser Pro 1 5 60 WO2007 108559 Al 2007 9 27 Ser Ser Leu Ala Ser Val Gly 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala 20 Leu Ala Trp Tyr Gln Gln Lys Pro 35 40 Tyr Asp Ala Ser Ser Leu Glu Ser 50 55 Ser Gly Ser Gly Thr Asp Phe Thr 65 70 Glu Asp Phe Ala Thr Tyr Tyr Cys 85 Thr Phe Gly Gln Gly Thr Lys Val 100 lt 210 gt 8 lt 211 gt 124 lt 212 gt PRT lt 213 gt Homo sapiens lt 400 gt 8 Glu Val Gln Leu Val Glu Cys Gl
169. Pro Gln Val Tyr Thr Leu 275 280 Thr Lys Asn Gln Val Ser Leu Thr Cys 290 295 Ser Asp Ile Ala Val Glu Trp Glu Ser 305 310 Tyr Lys Thr Thr Pro Pro Val Leu Asp 325 Tyr Ser Lys Leu Thr Val Asp Lys Ser 340 345 Phe Ser Cys Ser Val Met His Glu Ala 355 360 Lys Ser Leu Ser Leu Ser Leu Gly Lys 370 375 92 02007 108559 Al 2007 9 27 Val Phe Leu Phe Pro Pro Lys 170 175 Thr Pro Glu Val Thr Cys Val 190 Glu Val Gln Phe Asn Trp Tyr 205 Lys Thr Lys Pro Arg Glu Glu 220 Ser Val Leu Thr Val Leu His 235 240 Lys Cys Lys Val Ser Asn Lys 250 255 Ile Ser Lys Ala Lys Gly Gln 270 Pro Pro Ser Gln Glu Glu Met 285 Leu Val Lys Gly Phe Tyr Pro 300 Asn Gly Gln Pro Glu Asn Asn 315 320 Ser Asp Gly Ser Phe Phe Leu 330 335 Arg Trp Gln Glu Gly Asn Val 350 Leu His Asn His Tyr Thr Gln 365 10 20 30 40 93 WO2007 108559 Al 2007 9 27 lt 210 gt 76 lt 211 gt 377 lt 212 gt PRT 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence 64344 lt 400 gt 76 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 65 70 75 80 Tyr Thr Cys Asn Val A
170. Q 6 I 1 H S v A 9 S OLI 211 993 911 OLI LIL 961 DIL SHY 139 933 211 383 912 393 999 LUL 994 BOD HIL 331 339 288 331 JLH 33H JLL WOO 309 HH 919 121 389 939 1 1 399 JOU 199 19 ZZ A S 9 8 A A 3 1 9 3 0 d b A A H H V A q 4 48b je 996 BOO 199 989 228 212 099 933 211 200 100 119 220 949 JJH 010 281 993 991 118 199 319 991 OLI 299 JHH 999 BOO 139 993 218 991 918 239 161 LUO 149 111 ZSL 1 EI 9 5 v y 2 5 1 a 1 5 a 9 d b 1 9 9 9 S 422 991 199 999 109 998 989 121 98H DIL 999 H99 219 182 JHH 822 133 299 212 JLL 890 121 229 829 191 221 212 HOY 913 221 299 159 949 519 911 209 HOD 999 121 9b9 919 912 y 1 14 219 112 218 982 820 111 LHH 692 288 HUS 228 921 999 122 JHH 212 381 9919913983 982 919 LIL 9 319 100 HLL 199 OLL LLI JLL LIB 901 298 812 H99 DLL 989 81H 2289989319 L 4 vx 02007 108559 1 2007 9 27 116 1000 0 1000000 00 0 0 0 44099 06 GO x 8 QDWnHL O d A 90 LEES 6 7 40 er toarr o 80 Wu
171. Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205 His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser 210 215 220 Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 225 lt 210 gt lt 211 gt lt 212 gt lt 213 gt lt 220 gt lt 223 gt 230 235 85 107 PRT Artificial Sequence Description of Artificial Sequence 7 10VL_V104L mutant 10 20 30 40 107 WO2007 108559 Al 2007 9 27 lt 400 gt 85 Ala Gln Leu Thr Gln Ser Pro Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala 20 25 30 Leu Ala Trp Tyr Gln Gin Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 100 105 lt 210 gt 86 lt 211 gt 107 lt 212 gt PRT lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence 7 10VL_G100Q mutant lt 400 gt 86 Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 10 20 30 40 108 02007 108559 Al 2007 9 27 Asp Arg Val Thr Thr Cys Arg Ala Ser Gin Gly Ser Ser Ala 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu L
172. Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 lt 210 gt 4 lt 211 gt 124 lt 212 gt PRT lt 213 gt Homo sapiens lt 400 gt 4 Glu Glu Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 10 20 30 40 58 WO2007 108559 Al 2007 9 27 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Asp 20 25 30 Ala Met Tyr Trp Val Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ser Trp Asn Ser Gly Ser Gly Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Lys Ala Leu Trp Phe Gly Glu Phe Pro His Tyr Tyr Gly Met Asp 100 105 110 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 lt 210 gt 5 lt 211 gt 108 lt 212 gt PRT lt 213 gt Homo sapiens lt 400 gt 5 Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Thr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 10 20 30 40 59 Glu Asp Phe Ala Thr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro 85 90 Thr
173. cacaaagtct ttcaacaggg tgctctggct ctgcatctgt ctttagcctg ccagtttgga ctctcaccat atagttaccc caccatctgt ttgtgtgcct acgccctcca cctacagcct acgcctgcga cccaggtgcc aggagacaga gtatcagcag aagtggggtc cagcagcctg gctcactttc cttcatcttc gctgaataac atcgggtaac cagcagcacc agtcacccat gagagtgttg a 223 Description of Artificial Sequence 7 10G4344uhm L chain 400 84 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Pro Gly Ala Arg Cys Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser 20 25 30 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45 Gln Gly Ile Ser Ser Ala Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60 60 120 180 240 300 360 420 480 540 600 660 711 10 20 30 40 Ala Pro 65 106 WO2007 108559 1 2007 9 27 Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val 70 75 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Tle Ser Phe Asn Lys Arg 130 Glu Gln 145 85 90 95 Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 100 105 110 Ser Tyr Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 115 120 125 Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 135 140 Leu Lys Ser Gly Thr Ala Ser Val Va Cys Leu Leu Asn Asn 150 155 160 Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 165 170 175 Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp 180 185 190 Ser
174. hk 20 5 GTT GCT TGT GGG 3 00000 190 100 1000000000000000 94 30 72 C 30 000000050 000 94 30 70 C 300 72 C 30 000000050000 94 C 30 68 C 72 C 30 0000000 25000000 21 IO Tricine EDTA Buffer 98u 10000000000 5100 PER Nested sterile HO 30 PCR O0 0 O0 500 000 5 KOD PI us buffer 10X 5 dNTP Mx 2nM MSO 25 nM 21 KOD Pl us 1 unit ul Nested Universal primer NUP 100 1p Gene specific primers GSP 10 lu Total volume 50 100 10000000000000000000 00000 SM ART RACE cDNA 62000000 4 490 6 4 50 6 5 20 0 000 962 134 7 10 50 100 94000000100 000 94 C 50 H H 6 8 C 10 72 C 3 100000 200000008 72 C 7 000000 Ricca 154 02007 108559 Al 2007 9 27
175. tatgccatgt agttggaata agagacaacg gccttgtatt tggggccaag 125 taaaaggtgt gcaggtccct actgggtccg gtggtgacat ccaagaactc actgtgcaag ggaccacggt ccagtgtgaa gagactctcc gcaagctcca aggctatgcg cctgtatctg ggatgcgggg caccgtctcc 60 120 180 240 300 360 420 423 30 40 73 WO2007 108559 Al 2007 9 27 210 gt 32 lt 211 gt 141 lt 212 gt 213 gt sapiens lt 400 gt 32 Met Glu Leu Gly Leu Ser Trp Phe Leu Leu Ala Leu Lys Gly 1 5 10 15 10 Val Gln Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln 20 25 30 Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe 35 40 45 Asp Asn Tyr Ala Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 20 Glu Trp Val Ser Gly Ile Ser Trp Asn Ser Gly Asp Ile Gly Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn 85 90 95 Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu 100 105 110 30 Tyr Tyr Cys Ala Arg Asp Ala Gly Phe Gly Glu Phe His Tyr Gly Leu 115 120 125 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 130 135 140 lt 210 gt 33 9 lt 211 gt 390 lt 212 gt DNA 213 gt sapiens lt 400 gt 33 atggacatga gggtccccgc tcagctcctg gggcttctgc tgctctggct cccaggtgcc 60 agatgtgcca tccagttgac ccagtctcca tcctccctgt ctgcatctgt aggagacaga 120 74 02007 108559 Al 2007 9 27 gtcaccatca cttgccgggc aagtcagggc attagcagtg ctttagcctg gtatcagca
176. 0 2 11 100000000000000 60100000000 G4 Kabat 228H 2 350 000 1 8 0 125 WO2007 108559 1 2007 9 1 0 2 3 4 5 6 7 8 1000010000000 1000000000000 HHHHHHHHHHHHHHHH 00000000 10000000000000000 1 00000 2 10000
177. 0000 D000 00000000000000000000 O00000 00000 100000000 100000 0 1000000000 UOUO0O0O0000000000000000000000000000000000 UOUO0O0O0U000000000000000000000000000000000 100 O 100 HvHHHHHHHH UOUO0O0O0000000000000000000000000000000000 1000000000 1000000 0 1000000 1000000000 10000000 100000 IlHHUHHkHHHHH UOUO0O0O00000YyO00o00000000000000000000000000 10 20 30 40 50 39 02007 108559 1 2007 9 27 noon 10 20 30 40 50 40 WO2007 108559 Al 2007 9 27 H 10
178. 00000000000000000000000000000 Rs ZO EE ELTE 2 ep a a a tT um 2 4 79 0000000000000000000000000000000 Orita et al Blood 200 5 Jan 15 105 2 562 6 80000000 0 0 0 0 0 0 00 0 0 0 0 0 0 0 0 0 0 0 00 000 1000000000000000000000000000000 000000000000000000000000000000 n L1 1
179. 006 3 14 9 FERM BP 10558 2006 3 14 J 1 J 10 20 30 40 50 23 WO2007 108559 1 2007 9 27 10 20 30 40 50 WO2007 108559 Al 2007 9 27
180. 01 i A61K39 395 2006 01 i A61P7 00 2006 01 1 A61P7 04 2006 01 i A61P43 00 2006 01 i COTK16 28 2006 01 1 CO7K16 46 2006 01 1 C12N15 02 2006 01 i C12P21 08 2006 01 i I PC C12N15 00 15 90 CO7K16 00 16 46 JSTPlus JDream2 JMEDPlus JDream2 BIOSIS MEDLINE WPIDS STN GenBank EMBL DDBJ GeneSeq SwissProt PIR Geneseq DENG Bijia et al An agonist murine monoclonal antibody to the human c Mpl receptor stimulates megakaryocytopoiesis Blood 1998 Vol 92 No 6 pages 1981 1988 DEBILI Najet et al The Mpl ligand or thrombopoietin or megakaryocyte growth and differentiative factor has both direct proliferative and differentiative activities on human megakaryocyte progenitors Blood 1995 Vol 86 No 7 pages 2516 2525 C Di
181. 1 gt 29 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 64 gaactcaggt gctgggcacc ttgggcacg 210 65 lt 211 gt 33 212 DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 65 ccaaggtgcc cagcacctga gttcgagggg gga WO2007 108559 Al 2007 9 27 30 21 29 33 10 20 30 40 87 lt 210 gt 66 lt 211 gt 33 lt 212 gt DNA lt 213 gt Artificial lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 66 agagagggat cctcatttac ccagagacag gga lt 210 gt 67 lt 211 gt 27 212 DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 67 agagaggcta gcaccaaggg gccatcc 210 68 211 38 212 DNA lt 213 gt Artificial Sequence 220 223 Description of Artificial Sequence primer lt 400 gt 68 ggttttgagc tcaactctct tgtccacctt ggtgttgc lt 210 gt 69 lt 211 gt 40 212 DNA lt 213 gt Artificial Sequence lt 220 gt WO2007 108559 Al 2007 9 27 33 21 38 10 20 30 40 88 lt 223 gt Description of Artificial Sequence primer lt 400 gt 69 gtggacaaga gagttgagct caaaacccca cttggtgaca lt 210 gt 70 lt 211 gt 33 212 gt DNA lt 213 gt Artificial Sequence lt
182. 1_F 5 GCACCAAGGG CCCATCG 3 63 G3G4 P2 R 5 GAACTCAGGT GCTGGGCACC TTGGGCACG 3 64 G3G4_P3_F 5 CCAAGGTGCC CAGCACCTGA GTTCGAGGGG GGA 3 65 G3G4 P4 R 5 AGAGAGGGAT CCTCATTTAC CCAGAGACAG GGA 3 66 G434_P5_F 5 AGAGAGGCTA GCACCAAGGG GCCATCC 3 67 6434 5 GGTTTTGAGC TGTCCACCTT GGTGTTGC 3 68 G434_P7_F 5 GTGGACAAGA GAGTTGAGCT CAAAACCCCA CTTGGTGACA 3 69
183. 2 383 ILL 991 999 131 H32 933 DLL 21H 211 319 121 H32 199 919 DOH 192 HHH JLU 919 HH 398 999 ELE 9 9 4 1 1 S N s 0 0 9 A A 1 9 3 122 922 209 918 ULL 219 115 OLY 199 112 806 399 ILL 199 212 999 199 LUY 111 949 BHD LOL OBL 181 LIB 690 111 189 190 912 b 165 I 1 1 1 4 9 5 0 S 9 S S dle 219 921 891 081 901 HH HLI 191 222 122 932 921 998 221 188 199 982 913 398 318 998 212 129 211 198 599 LOL 699 196 299 298 211 908 BIL UID ESZ 8 6 301 5 5 q A l 11 3 9 d A 049 992 332 HIL 119 999 HIL 999 ULI JHL 389 JLL U99 LLL 999 199 111 219 319 999 19H BB9 911 199 221 229 1U9 JLU 919 212 988 122 159 999 599 983 06 wv 1 S 5 I 9 8 v L 1 A A 9462 210 208 999 LUY 409 931 999 JLO BIL 192 990 938 K L 991 949 12 919 963 091 329 ULL 129 198 208 118 299 969 LOU BOB 993 391 138 ILY 338 219 QUO 221 8 4 S v S 1 5 S S b 1 8 wL3 yw 9 139 189 193 998 190 219 IHY 219 OHL 909 LOL 993 822 199 B99 121 HID 121 913 221 221 922 LOL 989 908 9 1 983 21H 399 191 329 199 99 9 A T 1 1 T 98 1 1 0 228 9H9 289 HHO 222 289 080 210 599 982 221 IHL ALI 381 901089898121 913
184. 200 Asp Gly Val Glu Val His Asn Ala Lys 125 Pro Pro Cys Pro Arg 140 Pro Cys Pro Arg Cys 155 Phe Leu Phe Pro Pro 170 Pro Glu Val Thr Cys 190 Val Gln Phe Lys Trp 205 Thr Lys Leu Arg Glu Cys Pro Pro Ala 160 Lys Pro 175 Val Val Tyr Val Glu Gln 10 20 30 40 210 215 Asn Ser Thr Phe Arg Val 225 230 Asp Trp Leu Asn Gly Lys Glu 245 Leu Pro Ala Pro Glu Lys 260 Arg Glu Pro Gln Val Tyr Thr 275 Lys Asn Gln Val Ser Leu Thr 290 295 Asp Ile Ala Val Glu Trp Glu 305 310 Asn Thr Thr Pro Pro Met Leu 325 Ser Lys Leu Thr Val Asp Lys 340 Ser Cys Ser Val Met His Glu 355 Ser Leu Ser Leu Ser Pro Gly 370 375 lt 210 gt 56 lt 211 gt 1199 lt 212 gt DNA lt 213 gt Homo sapiens lt 400 gt 56 83 WO2007 108559 Al 2007 9 27 220 Val Ser Val Leu Thr Val Leu 235 Tyr Lys Cys Lys Val Ser Asn 250 Thr Ile Ser Lys Thr Lys Gly 265 270 Leu Pro Pro Ser Arg Glu Glu 280 285 Cys Leu Val Lys Gly Phe Tyr 300 Ser Asn Gly Gln Pro Glu Asn 315 Asp Ser Asp Gly Ser Phe Phe 330 Ser Arg Trp Gln Gln Gly Asn 345 350 Ala Leu His Asn Arg Tyr Thr 360 365 Lys His Gln 240 Lys Ala 205 Gln Pro Met Thr Pro Ser Asn Tyr 320 Leu Tyr 335 Tle Phe Gln Lys 10 20 30 40 ctagcaccaa gggcccatcg gtcttccccc tggcgccctg ctccaggagc acctctgggg 60 gcacagcggc cctgggctgc ctggtcaagg actacttccc cgaaccggtg acggtgtcgt 120 ggaactca
185. 2007 9 27 10 BS 5ng PEG r 1 50m0 0000 00 00000 0 1 500rpm 5mn 4 CH 0 1096851 0 IMDM
186. 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 70 aggtgctggg caccgtgggc atgtgtgagt tgt lt 210 gt 71 lt 211 gt 33 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 71 cacacatgcc cacggtgccc agcacctgag ttc lt 210 gt 72 lt 211 gt 57 lt 212 gt lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 72 WO2007 108559 Al 2007 9 27 40 33 33 tgtgtgagtt gtgtcaccaa gtggggtttt ggactcaact ctcttgtcca ccttggt 57 10 20 30 40 89 WO2007 108559 Al 2007 9 27 lt 210 gt 73 lt 211 gt 57 lt 212 gt lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 73 accccacttg gtgacacaac tcacacatgc ccaccatgcc cagcacctga gttcgag lt 210 gt 74 lt 211 gt 347 lt 212 gt PRT lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence 6334451 lt 400 gt 74 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr 65 70
187. 288 3550 s 5585 u 5585 358 D 3 3 3 d 3 1 4 V N H A 3 A 8 Q0 A A A N 4 A 3 d 86 ma ma 22222120 25935 za 219 213 212 229 399 211 191 211 999 BIS 280 212 282 999 ULI 383 OLY 228 011 919 983 212 995 88323239 SBSSSSS BBSSSSS 85932222 SBSSSSSS BBSSSSS SSSSSSS 982 993 999 BOY JHH 339 LUY 182 919 919 399 140 919 JHL 991 ILL 983 219 222 296 evo C do c lt q 3 0 8 A A A 3 1 3 d L 8 S N 1 L q 3 d 812 112 948 531 383 813 383 383 92H 291 983 943 659 951 399 908 9 1 IHL HOL 513 311 999 LIL DUD 20H 919 940 919 O10 610 301 52H 210 OUO 122 308 993 DIL 21H 91H 212 126 280 9bH 222 228 d d 3 1 4 A 8 d 9 9 3 3 V d 3 L H 1 199 999 OY 949 199 199 123 209 212 946 212 HOO 199 199 938 199 199 SIY LOL 919 YIL L L 919 hes 1adN BOJ 222 311 OLI 211 319 BOO UNO 999 211 9b9 159 499 HII 201 BOO 899 391 2383 12H 389 252 9 1 d L M 8 H A M LN S d X3 Hd A N 3 1 4 zz TEBE 899 bb9 199 991 LLL 906 212 BBO 191 ILL 912 292 ILL 991 ILL 901 999 211 919 HLI LUI 911 92H 991 919 amp FIORI 188 112 BOO 22H HHH 231 LLO 288 O10 OHU 208 JHH 208 200 OW 280 180 HLO JHH JOL 338 391 229 c uu wWw
188. 3 4 MDS 5 1000000 6 00000000 1000000 FE ELEC EHE SER CER EPI 0 0 10000000000000000 126 02007 108559 Al 2007 9 27 DI HHHHHHHHHHHHHHHHHHHHHHc MI LJ 1 1000000000000000 Thrombopoi eti TPO 0 0 332 1 9 N HHHHHHHHHHHHHHHHHHHHHHHH 4 79 9 000000000 63558 00000000 6100 00000000000000000000000 1000000000000000000000000 00000000 1000000 Genbank NM 005373 _0053641 000
189. 3333 0000000 2222222 M 4 1 1 S A 0 N M LU 3 3 b S d d UD 445 5054502 5545485 2522222 SBEESEE 5555555 LLL 983 02H 991 280 931 983 919 OLL 211 991 381 212 212 219 998 190 000 001 944 219 199 EE EE 2222209 2522202 219 913 291 338 912 29H 219 982 HU 22H ALY 983 221 495 222 949 208 394 919 949 HII 2211 33533335 022222 IERI 2525255 2258555 3 d 0 9 X v S 1 X 3 S S d 1 9 X N S A X 2 NE EEEEEEE 6555 Sezze 5555555 212 128 999 219 222 111 999 lil 998 991 111 212 981 998 998 399 990 922 111 011 998 983 311 939 arannana snansoona ZEEE EEEEEEE 989 UNI 222 983 999 HUU 229 221 218 009 218 221 221 999 212 299 221 219 288 291 2581 4424444 EEEE 53202222 PC Oh H N 3 3 9 N 1 A d 6 H 1 A S 4 3 1 S N 555656655 2555255 EE 52 JLL 918 213 DLL 999 911 2HO 338 913 219 819 289 940 921 983 282 828 91H 291 931 911 986 bold MN UC MR E 3433313 99595959 9555959 SHO 299 913 981 982 383 912 218 338 212 219 398 348 919 199 281 938 JHH 211 46 3
190. 7 1964344 000000 1gG4344 HHHHHHHN5KG3331H 0000000000000 643 4 P5 6434 P6 94 1000101000000 94 150 55 C 109 68 C I00000000320000000072Cc700000000000N2kKG 4PE O O G434 7 GG P2 RIH I I D DU 0O O PCR j l DNA 0 0 Q Aqui ck Gel Extraction K it DNA N5KG4PEQ G3G4 6364 P4 100000000000 DNA OOO 30 Overlap Extenti on DNAI 100 94 0 1 0 O 94 C 10 55 C 100 68 C 1 5 100 5 1000000000000000 434 P5 PA RI 94 C 50 68 2 2000 72 C 7U7 00000 PCRI QAquick Extraction Kit 00000 pCR 4 Blunt 1000000 100 DN J JUD U U U 1000000 19643441 0000000000000000 P5 5 AGAGAGGCTA GCACCAAGGG GCCATCC 3 O 670 0434 P6 5 GGITITGAGC TGTCCACCTT GGTGTTGC 3 68H P7 5 GTGGACAAGA GAGTTGAGCT CAAAACCCCA 3 00000 690 100000 8 196434401 1000 163 02007 108559 1 2007 9 27
191. 900060 14 5 20 Uu U KCo5 j DO 19 00000000 140 5820 000000 Tom zuka et al Proc Natl Acad Sci USA 2000 Vol 97 72207 00000 B lgH HHkHHHHHHHHHHHHHHHHHHHHHHl9 KCo5 KM HHHHHHHHHHHHHHHHIgHHHHHk 0000 0010 Nat Biotechnol 1996 Vol 14 845 WOO L L3 L3 L3 D r O 137 02007 108559 1 2007 9 27
192. AGTCTGGGGGAGGCTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCTTT GATGATTATGCCATGTACTGGGTCCGGCAAGTTCCAGGGAAGGGCCTGGAGTGGGTCTCAGGTATTAGTTGG AACAGTGGTAGCATAGGCTATGCGGACTCTGTGAAGGGCCGATTCACCGTTTCCAGAGACAACGCCAAGAAC TCCCTGTATCTGCAAATGAACAGTCTGAGAGCTGAGGACACGGCCTTATATTACTGTGCAAAAGCCCTATGG TTCGGGGAGTTCCCCCACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA 27 00 000000000000000000000000000 E MELGLSWIFLVAILKGVQCEEQLVESGGGLVQPGRSLRLSCTASGFTFDDYAMYWVRQVPGKGLEWVSGISW NSGSIGYADSVKGRFTVSRDNAKNSLYLQMNSLRAEDTALYYCAKALWFGEFPHYYGMDVWGQGTTVTVSS 28 U0O0 0000000000000000000000000000000000000 IIHHHHHHHHH ATGGACATGAGGGTCCCCGCTCAGCTCCTGGGGCTTCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCATC CAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGT CAGGGCATTAGCAGTACTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGAT GCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACC ATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCCGTACACTTTT GGCCAGGGGACCAAGCTGGAGATCAAACGT 29 00 000000000000000000000000000 I0O000000000000000000000000000000 MDMRVPAQLLGLLLLWLPGARCAIQLTQSPSSLSASVGDRVTITCRASQGISSTLAWYQQKPGKAPKLLIYD
193. CACACATGCCCACGGTGCCCAGAGCCCAAATCTTGTG ACACACCTCCCCCGTGCCCACGGTGCCCAGAGCCCAAATCTTGTGACACACCTCCCCCATGCCCACGGTGCC CAGAGCCCAAATCTTGTGACACACCTCCCCCGTGCCCAAGGTGCCCAGCACCTGAACTCCTGGGAGGACCGT CAGTCTTCCTCTTCCCCCCAAAACCCAAGGATACCCTTATGATTTCCCGGACCCCTGAGGTCACGTGCGTGG TGGTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAAGTGGTACGTGGACGGCGTGGAGGTGCATAATG CCAAGACAAAGCTGCGGGAGGAGCAGTACAACAGCACGTTCCGTGTGGTCAGCGTCCTCACCGTCCTGCACC AGGACTGGCTGAACGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAA CCATCTCCAAAACCAAAGGACAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGA CCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGCGACATCGCCGTGGAGTGGGAGA GCAATGGGCAGCCGGAGAACAACTACAACACCACGCCTCCCATGCTGGACTCCGACGGCTCCTTCTTCCTCT ACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACATCTTCTCATGCTCCGTGATGCATGAGG CTCTGCACAACCGCTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATGA 56 HHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH HHHH HHHHCHHHHH CHHHHHHCHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH 000000 lt 000000000000000000000000000 0 10000000000 1000000000000000000000000000 00000 000000 CHHHHHHHHHHHHHHHHHHHHHHHH 0 000000000000000000000000000000000000000 VA EE EI BE CHE E A 00000000000 45 02007 108559 1 2007 9 27
194. CCATGGACATGAGGGTCCCCGCTC 3 43 LV 3 165_1B_L18_Bsi 5 AGAGAGAGAG CGTACGTTTG ATCTCCACCT TGGTCCCTCC 3 44 4 49 10 LV 5 DNP_LIBglp 5 AGAGAGAGAGATCTCTCACCATGAGGGTCCCCGCTCAGCTC 3 45 LV 3 A27_R_N202 5 AGAGAGAGAGCGTACGTTTGATTTCCACCTTGGTCCCTTGGC 3 46 E C gt E E HHHHHHHHHHHHHHHHHH HHHHHHHHHHHHHHHHHHHH 20 30 40 42 02007 108559 Al 2007 9 27 6 4 50 LV 5 208LF 5 AGAGAGAGAGATCTCTCACCATGGACATGAGGGTCCCCGCTCAGC 3 47 LV 3 62LP3Bsi 5 AGAGAGAGAGCGTACGTTTGATTTCCACCTTGGTCCCTTG 3 48 6 5 2 35 LV 5
195. CCT GCCCCCAT CCCGGGAGGAGAT GACCAAGAACCAGGT CAG CCT GACCT GCCT GGT CAAAGGCTT CT ACCCCAGCGACAT CGCCGT GGAGT GGGAGAGCAAT GGGCAGCCGGAGAACAACT ACAACACCACGCCT CCCAT GGACT CCGACGGCT CCTTCTT CCT CT ACAGCAAGCT CACCGT GGACAAGAGCAGGT GG CAGCAGGGGAACATCTTCT CAT GCT CCGT GAT GCATGAGGCT CT GCACAACCGCTACACGCAGAAGAGCCT CT CCCTGTC TCCGGGTAAATGA DO 0 0 560 100000 4 1963311100000 9633110 0 NSKG3H 00000000000 linkH 1
196. CTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGT CAGGGCATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATGAT GCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACC ATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCCGTGGACGTTC 30 GGCCAAGGGACCAAGGTGGAAATCAAACGT 33 10 0 000000000000000000000000000 ee OA AO MDMRVPAQLLGLLLLWLPGARCAIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKVPKLLIYD ASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPWTFGQGTKVEIKR 34 UOU0O0000 0 0000000000000000000000000000 IO0O0U0O0000000000000000000000000000000000 OA A HHHHHHHHHHH 40 38 02007 108559 1 2007 9 27 ATGGAGTTGGGACTGAGCTGGATTTTCCTTTTGGCTATTTTAAAAGGTGTCCAGTGTGAAGTGCAACTGGTG GAGTGTGGGGGAGGCTTGGTACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTT GATGATTATGCCATGCACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGGAGTGGGTCTCAGGTATTAGTTGG AATAGTGGTAGTATAGGTTATGCGGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAAC TCCCTGTATCTGCAAATGAACAGTCTGAGAGCTGAGGACACGGCCTTGTATTACTGTGCAAAACCTATATGG TTCGGGGAGTGGGGAAACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA AE 35 HH H HHHHHHHHHHHHHHHHHHHHHHHHHH
197. EEE 3553555353 20500950 90H 991 211 219 LOL 918 919 OLI 949 212 JHL 90H 198 DLL SSSSSSS ggg 5555555 EEEEEEE 812 131 913 901 212 298 982 282 919 129 989 LHI 944 919 301 391 VIL 211 319 2521 E E E E E EZEEZZE EE BE 5 E E E EEREEESE 9 3 0 A q A L 5 A 1 4 4 S 9 q S q d t 58552282252 883853538 2222222 239 212 219 398 221 921 211 OLI 269 991 199 001 991 918 HHL 022 919 998 912 389 282 999 3555535 4433448 999 991 99H 288 949 228 HLI 99H 398 381 313 ILL ILL 221 299 389 221 389 812 919 220 2221 BORAZZA d 1 1 A N N 3 q 0 9 N S 3 A 3 A NV d A 4 9 486 da gd JAAR 555555 11 B99 291 901 211 OLY 911 9LL 212 969 919 999 ULL 994 212 338 212 383 999 JHL OLI 921 999 O1H 922 Ppp pap EZEEEEE 122 93H 999 909 1BH 29H 061 940 019 229 218 289 398 222 281 ILL 300 2921 TITI RTRT EE 383
198. GTLELRPRSRYRLQLRARLNGPTYQGPWSSWSDPTRVETATETAW COOH Ad 17 x 10 20 30 40 50 30 WO2007 108559 1 2007 9 27 nnm E PIE nn E x D D D HH H C m n n n C n n DIE
199. HHHHHHHH HH 000012 100 HH 7 10G3311 4 49G3311H D 0 0 10000000 APO 0 0000000000000000000 DO HHHHHHHHHHHHHHHHHHHHHHHH 13 DO PEG HuMGDF 0 HHHHHHHHHHHHHH NOoGHHHHHHHHHH CD34 HHH1 000HHHHH 10 0000 0 IHHHHHHHHHHHHHHHHHHHHHHHHHHHHH PreHHHHHHHHH NoGHHHHHHHHHH CD34 HHH1 000HHHHH 10 000HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH 171 02007 108559 1 2007 9 27
200. HHHHHHHHHH 0 0 100000000000000000000000000000 101000 00000000 IIH 100000 7 10G4344uhnf gGAPE O 100000000000000 HLA DR DQ DPO Al cs 100000 100000000000000 1000 Mol D 00 00000 10000000000 0000000 10000000000 5 5kb PCR pBluescript 0 13 000000 Mpl 0 0000 78 600 0 10000000000000000000000000000 DNAD 0 O PRO TO 0000000000000 199 00000 557860000 0000000 0000000000 39000000 0000000000 RT PCRO OO 0 390 0000000000000 7 10G4344uhn 000003 10 gm0 000000000000 KX 21H 10000000000000000 100000000 0 0 0 PEGrHuMGDF 100000 7 10G4344uhm10u g 11 7 10G4344uhm 9 gl 00 IV PBS 169 02007 108559 Al 2007 9 27
201. HHHHHHHHHHHHH N CH1 CH2 CH3H 0 0 0 0 0 1000000000000000000000 CHIH 0800000000 0 19630 000000 H2 cH30 0 O0 196 0000000000000000000000000000 CH1 CH2 O OO 00 0 0 0 0 00 0 1963 3 711 19G3311 7 1000000000000000000000 903 CH2H 19G4PEH 100000000 1943440 00000 963 100000 0 0 uppe 94 00000000000000000000000 Kabat EU ngU Kabat Sequences of Proteins of Immunological Interest 5thEd Pub Health Service National Institute of Health Bethesda 1991 000 2160 226 2260 231 1000000000000000000000000 5 15 3 0000000000 196300000 100000000000000000 635100000 00000 lgGx3xxh1 x PM nnnnnm 0000 1963 0000000000000000000000000 uppe mddle H Kabat EU Sequences of Proteins of Immunological Interest 5thEd Pub Health Service National Institute of Health Bethesda 1991
202. HHHHHHHHHHHHHHHHH UOUO0O0O0U00000000000000000000000000000050 U0O0 00u00000000000000000000000000000000050 1000000000 E 1000000000 ATGGACATGAGGGTCCCCGCTCAGCTCCTGGGGCTTCTGCTGCTCTGGCTCCCAGGTGCCAGATGTGCCATC CAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGT CAGGGCATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGAT GCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACC ATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCCGCTCACTTTC GGCGGAGGGACCAAGGTGGAGATCAAA 25 IO0 000000000000000000000000000 IO0OU000000000000000000000000000050 MELGLSWIFLLAILKGVQCEVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAMHWVRQAPGKGLEWVSGISW NSGSIGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALY YCAKNLWFGEFRYWYFDLWGRGTLVTVSS 26 UOUO0O0000 00000000000000000000000000000 1000000000000000000000000000000000000 00 000000000000000000000000000000000000 10 20 30 40 50 36 WO2007 108559 Al 2007 9 27 sp ESE OG E ATGGAGTTGGGACTGAGCTGGATTTTCCTTGTGGCTATTTTAAAAGGTGTCCAGTGTGAAGAGCAGCTGGTG G
203. Leu Gly Lys 370 375 10 20 30 40 95 WO2007 108559 Al 2007 9 27 lt 210 gt 77 lt 211 gt 347 lt 212 gt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence G4344hl lt 400 gt 77 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 65 70 75 80 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val Glu Leu Lys Thr Pro Leu Gly Asp Thr Thr His Thr Cys Pro 100 105 110 Arg Cys Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg 115 120 125 Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe Leu Phe Pro 130 135 140 Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr 145 150 155 160 Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn 165 170 175 10 20 30 40 1 Glu Glu Gln 195 Leu His Gln 210 Asn Lys Gly 225 Gly Gln Pro Glu Met Thr Tyr Pro Ser 275 Asn Asn Tyr 290 Phe Leu Tyr 305 Asn Val Phe Thr Gln Lys lt 210 gt 78 lt 211 gt 257 lt 212 gt PRT 96 WO2007 108559 Al 2007 9 27 Asp Gly Val Glu Val His Asn Ala Ly
204. Pro Ser Val Phe Leu Phe 10 20 30 40 99 02007 108559 Al 2007 9 27 115 120 125 Pro Pro Lys Pro Lys Asp Thr Leu Met Ser Arg Thr Pro Glu Val 130 135 140 Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe 145 150 155 160 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 165 170 175 Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr 180 185 190 Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 195 200 205 Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala 210 215 220 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln 225 230 235 240 Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 245 250 255 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 260 265 270 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser 275 280 285 Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu 290 295 300 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 305 310 315 320 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys 325 330 10 20 30 40 100 210 gt 80 211 gt 327 212 gt PRT 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence G4PE lt 400 gt 80 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 1 5 10 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu 20 25 Phe Pro Glu Pro Val Thr Val Ser Trp Asn S
205. T CCT GGGGCTT CT GCT GCT CT GGCT CCCAGGT GCCAGAT GT GCCAT CCAGTT GAC CCAGTCTCCAT CCT CCCT GT CT GCAT CT GTAGGAGACAGAGT CACCAT CACTT GCCGGGCAAGT CAGGGCATT AGCAGT G gt ATG 0000000000000 DNA 1000 156 02007 108559 1 2007 9 27 CTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTC CCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAAC TTATTACTGTCAACAGTTTAATAGTTACCCGTGGACGTTCGGCCAAGGGACCAAGGTGGAAATCAAACGT lt 6 4 500 0000000 0000000000000000 10000000000000000000000000000000 MDMRVPAQLLGLLLLWLPGARCAI QLTQSPSSLSASVGDRVTI TCRASQGI SSALAVYOOKPGKVPKLLI YDASSLESGV PSRFSGSGSGTDFTLTI SSLQPEDFATYYCQQOF NSYPW FGOGTKVEI KRO 340 1000000 6 5 21000000000000000000000 94 00000 JDDUUDUHIEO
206. T I TCRASQGISSALAWYQQKPGKAPKLLI YDASSLESGVPSRFSGSGSGTDFT LTISSLQPEDFATYYCQQFNSYPLTFGGGTKVETK 7 10VL_V104L 4 49VL 85 AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFT LTISSLQPEDFATYYCQQFNSYPLTFGGGTKLEIK 7 10VL_G100Q 6 4 50VL 1 86 AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFT LTISSLQPEDFATYYCQQFNSYPLTFGQGTKVEIK 7 10VL A43V 6 4 50VL 2 87 AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKVPKLLIYDASSLESGVPSRFSGSGSGTDF TLTISSLQPEDFATYYCQQFNSYPLTFGGGTKVEIK 7 10VL_Y94F CDR 88 AIQLTQSPSSLSASVGDRVT I TCRASQGISSALAWYQQKPGKAPKLL I YDASSLESGVPSRFSGSGSGTDFT LTISSLQPEDFATY YCQQFNSFPLTFGGGTKVEIK oanad O nnnnnnnnnnnnnnnnnnnnnn 54 02007 108559 Al 2007 9 27
207. TGTACACC 0 0 O 620 1983344 196334411 gG4344 196434411 gG4344uh 1 gG4344uhm 10000000000000 HHHHHHHHHHHHHHHHHHH N5KG1 7 100 1961 L3 L3 L3 162 02007 108559 Al 2007 9 27
208. VGDRVTI TCRASQG SSALAW QOKPGKVPKLLI YDASSLESGVPSRF SGSGSGTDFTLTI SSLQP EDF ATYYCQOF NSYPLTF GGGTKVEI 7 10VL Y94F CDR 0 0 00 00 880 D Al QLTQSPSSLSASVGDRVT I TCRASQG SSALAW QOKPGKAPKLLI YDASSLESGVPSRF SGSGSGT DFTLTI SSLQP EDF ATYY CQQF NSF GGGTKVEI HHHHHHHHHH 1 0 1 0 01u FMBA hMplH HHHHHHHH HHHHHHHHHHHHHHH 1964 000000000000000 7 10G4344uhm 11 0000000000 0 7 000000000000 7 10643446 1200 HHHHHHHHHHHcMIHH 0001000 000000000 HHHHHHH HHHHHHHHHHHHHHHHH 10000000000000
209. WO2007 108559 Al 2007 9 27
210. acticable search terms used JSTPlus 2 JMEDPlus JDream2 Igaku Yakugaku Yokoshu Zenbun Database BIOSIS MEDLINE WPIDS STN GenBank EMBL DDBJ GeneSeq DOCUMENTS CONSIDERED TO BE RELEVANT Category Citation of document with indication where appropriate of the relevant passages Relevant to claim No DENG Bijia et al An agonist murine 1 12 monoclonal antibody to the human c Mpl receptor stimulates megakaryocytopoiesis Blood 1998 Vol 92 No 6 pages 1981 1988 DEBILI Najet et al The Mpl ligand or thrombopoietin or megakaryocyte growth and differentiative factor both direct proliferative and differentiative activities on human megakaryocyte progenitors Blood 1995 Vol 86 No 7 pages 2516 2525 Kazuhiro MOTOKI et al Hito Senshokutai Mouse o Mochiita Hito Kotai Sakusei to sono Rinsho Oyo Igaku no Ayumi 13 November 2004 13 11 04 Vol 211 No 7 pages 733 to 736 ce Further documents are listed in the continuation of Box C patent D Further documents are listed in the continuation of Box C See patent amiamo annex Special categories of cited documents T later document published after the international filing date or priority document defining the general state of the art which is not considered to date and not in conflict with the application but cited to understand be of particular relevance the principle or theory underlying the invention
211. amp AU 2003 201322 B2 EAPCT ISA 210 2 2005 4 182 02007 108559 Al 2007 9 27 PCT JP2007 056536 Al 1 2 8 3 PCT17 2 a 1 2 PC 6 4 2 3 1 3 PC 13 1
212. ctcacgtcca ccaccacgca acccagacct gttgagctca aaatcttgtg cctcccccat aggtgcccag aaggataccc cacgaagacc aagacaaagc gtcctgcacc ctcccagccc gtgtacaccc ctggtcaaag gagaacaact agcaagctca atgcatgagg tga 240 300 360 420 480 540 600 660 720 780 840 900 960 1020 1080 1133 10 20 30 27 40 30 85 lt 210 gt 59 lt 211 gt 30 lt 212 gt lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 59 tgcgtggtgg tggacgtgag ccacgaagac lt 210 gt 60 lt 211 gt 23 lt 212 gt lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 60 tgatcatacg tagatatcac lt 210 gt 61 lt 211 gt 30 212 DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 61 ggtgtacacc tgtggctctc ggggctgccc lt 210 gt 62 lt 211 gt 30 lt 212 gt lt 213 gt Artificial Sequence lt 220 gt WO2007 108559 Al 2007 9 27 30 23 30 10 20 30 40 86 223 gt Description of Artificial Sequence primer lt 400 gt 62 gggcagcccc gagagccaca ggtgtacacc lt 210 gt 63 lt 211 gt 27 lt 212 gt lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 63 agagaggeta gcaccaaggg cccatcg lt 210 gt 64 lt 21
213. ctcc 120 tgtgcagcct ctggattcac ctttgatgat tatgccatgc actgggtccg gcaagctcca 180 gggaagggcc tggagtgggt ctcaggtatt agttggaata gtggtagcat aggctatgcg 240 gactctgtga agggccgatt caccatctcc agagacaacg ccaagaactc cctgtatctg 300 caaatgaaca gtctgagagc tgaggacacg gccttgtatt actgtgcaaa aaatctatgg 360 ttcggggagt tccgttactg gtacttcgat ctctggggcc gtggcaccct ggtcactgtc 420 tcctca 426 210 24 lt 211 gt 142 212 PRT lt 213 gt Homo sapiens 400 24 Met Glu Leu Gly Leu Ser Trp Ile Phe Leu Leu Ala Ile Leu Lys Gly 1 5 10 15 Val Gln Cys Glu Val Gln Leu 20 Pro Gly Arg Ser Leu Arg Leu 35 Asp Asp Tyr Ala Met His Trp 50 55 Glu Trp Val Ser Gly Ile Ser 65 70 Asp Ser Val Lys Gly Arg Phe 85 Ser Leu Tyr Leu Gln Met Asn 100 Tyr Tyr Cys Ala Lys Asn Leu 115 Val Glu Ser Gly Gly 25 30 Gly Phe 45 Ser Cys Ala Ala Ser 40 Val Arg Gln Ala Pro 60 Gly Lys Trp Asn Ser Gly Ser 75 Gly Leu Val Thr Phe Gly Leu Ile Gly Tyr Ala 80 Thr Ile Ser Arg Asp Asn Ala Lys Asn 90 Ser Leu Arg Ala Glu 105 Trp Phe Gly Glu Phe 120 125 95 Asp Thr Ala Leu 110 Arg Tyr Trp Tyr 10 20 30 40 69 02007 108559 Al 2007 9 27 Phe Asp Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser 130 135 140 lt 210 gt 25 lt 211 gt 387 lt 212 gt DNA lt 213 gt Homo sapiens lt 400 gt 25 atggacatga gggtccccgc tcagctcctg gggcttctgc tgctctggct cccaggtgcc 60 agatgtgcca tccagttgac ccagtctcca tcctccctgt c
214. ctggccagg ctcccaggct cctcatctat gatgcatcca gcagggccac tggcatccca gacaggttca gtggcagtgg gtctgggaca gacttcactc tcaccatcag cagactggag cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcaccgat caccttcggc caagggacac gactggagat taaacgt lt 210 gt 38 lt 211 gt 129 lt 212 gt PRT lt 213 gt Homo sapiens lt 400 gt 38 Met Glu Thr Pro Ala Gln Leu Leu Phe Leu Leu Leu Leu Trp Leu Pro 1 10 15 Asp Thr Thr Gly Glu Val Leu Thr Gln Ser Pro Gly Thr Leu Ser 20 25 30 Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser 35 40 45 Val Ser Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala 50 55 60 Pro Arg Leu Leu Ile Tyr Asp Ala Ser Ser Arg Ala Thr Gly Ile Pro 65 70 75 80 60 120 180 240 300 360 387 10 20 30 40 77 02007 108559 1 2007 9 27 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 85 90 95 Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr 100 105 110 Gly Ser Ser Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu 115 120 125 Arg lt 210 gt 39 lt 211 gt 45 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 39 agagagagag gtcgaccacc atggagttgg gactgagctg gattt lt 210 gt 40 lt 211 gt 40 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 40 agagagagag gctagctgag gagacagtga ccagggt
215. earlier application or patent but published on or after the international filing document of particular relevance the claimed invention cannot be date considered novel or cannot be considered to involve an inventive document which may throw doubts on priority claim s or which is step when the document is taken alone cited to establish the publication date of another citation or other document of particular relevance the claimed invention cannot be special reason as specified considered to involve an inventive step when the document is document referring to an oral disclosure use exhibition or other means combined with one or more other such documents such combination document published prior to the international filing date but later than the being obvious to a person skilled in the ant priority date claimed g document member of the same patent family Date of the actual completion of the international search Date of mailing of the international search report 08 June 2007 08 06 07 19 June 2007 19 06 07 Name and mailing address of the ISA Authorized officer Japanese Patent Office Facsimile No Telephone No Form PCT ISA 210 second sheet April 2005 176 02007 108559 Al 2007 9 27 INTERNATIONAL SEARCH REPORT International application No Tre Continuation DOCUMENTS CONSIDERED TO BE RELEVANT DOCUMENTS CONSIDERED TO BE RELEVANT Citation of document with indication where appropriate of the rel
216. equence having a specific mutation in the amino acid sequence wherein all of the amino acid sequences mentioned above appear to be only for a specific heavy chain or a specific light chain of an antibody However even though the common technical knowledge at the time of filing the present application is taken into the consideration it is quite difficult to say that an antibody can certainly bind specifically to specific antigen and exert a desired function or property merely by defining only a heavy chain and a light chain of the antibody Therefore the specific DNA comprisinganucleotide sequence encoding specific amino acid sequence selected from the group consisting of the amino acid sequence depicted in a specific SEQ ID number and an amino acid sequence having a specific mutation in the amino acid sequence recited in these claims does not shear any common chemical structure or function but has such an only common feature that it is a kind of DNA Since DNA is already known at the time of filing the present application the common feature among the inventions of claims 14 15 17 and 18 specific DNA comprising a nucleotide sequence encoding a specific amino acid sequence selected fromthegroupconsistingoftheaminoacid sequence depicted ina specific SEQ ID number and an amino acid sequence having a specific mutation in the amino acid sequence cannot be regarded as a special technical feature in the meaning within PCT Rule 13 2 Therefo
217. er Gly 35 40 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 65 70 75 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr 85 90 Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro 100 105 Glu Phe Glu Gly Gly Pro Ser Val Phe Leu Phe Pro 115 120 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr 130 135 140 Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn 145 150 155 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg 165 170 WO2007 108559 Al 2007 9 27 Pro Cys Val Lys 30 Ala Leu 45 Gly Leu Gly Thr Lys Val Cys Pro 110 Pro Lys 125 Cys Val Trp Tyr Glu Glu Ser Arg 15 Asp Tyr Thr Ser Tyr Ser Lys Thr 80 Asp Lys 95 Ala Pro Pro Lys Val Val Val Asp 160 Gln Phe 175 10 20 30 40 101 02007 108559 1 2007 9 27 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln 180 185 190 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu 195 200 205 Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 210 215 220 10 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys 225 230 235 240 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245 250 255 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 260 265 270 20 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 275 280 285 Arg Leu Thr Val Asp Lys Ser A
218. er Trp Ser Asp Pro 465 470 475 480 Thr Arg Val Glu Thr Ala Thr Glu Thr Ala Trp 485 490 lt 210 gt 18 lt 211 gt 31 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 18 tcttgtccac cttggtgttg ctgggcttgt g 210 19 211 26 lt 212 gt DNA 213 Artificial Sequence 220 223 Description of Artificial Sequence primer 400 19 gttgaagctc tttgtgacgg gcgagc 31 26 10 20 30 40 67 lt 210 gt 20 lt 211 gt 30 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt 223 gt Description of Artificial Sequence primer lt 400 gt 20 aggcacacaa cagaggcagt tccagatttc lt 210 gt 21 lt 211 gt 25 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 21 gctggagggc acggtcacca cgctg 210 22 lt 211 gt 31 212 DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 22 tgcacgccgc tggtcagggc gcctgagttc lt 210 gt 23 lt 211 gt 426 lt 212 gt DNA lt 213 gt Homo sapiens WO2007 108559 Al 2007 9 27 30 25 3 10 20 30 40 68 WO2007 108559 1 2007 9 27 lt 400 gt 23 atggagttgg gactgagctg gattttcctt ttggctattt taaaaggtgt ccagtgtgaa 60 gtgcagctgg tggagtctgg gggaggcttg gtacagcctg gcaggtccct gagact
219. ernational application that do not comply with the prescribed requirements to such an extent that no meaningful international search can be carried out specifically 3 C Claims Nos because they are dependent claims and are not drafted in accordance with the second and third sentences of Rule 6 4 Box No Observations where unity of invention is lacking Continuation of item 3 of first sheet This International Searching Authority found multiple inventions in this international application as follows Where a group of inventions is claimed in one and the same international application the requirement of unity of invention PCT Rule 13 1 shall be fulfilledonlywhenthere isatechnical relationship among those inventions involving one or more of the same or corresponding special technical features The expression special technical feature shallmeanthose technical features that define a contribution which each of the claimed inventions considered as a whole makes over the prior art Rule 13 2 The determination of unity of inventions shall be made without regard to whether a group of inventions are claimed in separate claims or as alternatives within a single claim continued to extra sheet 1 0 As all required additional search fees were timely paid by the applicant this international search report covers all searchable claims 2 0 As all searchable claims could be searched without effort justifying additional
220. eu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 15 80 Glu Asp Phe Ala Thr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 lt 210 gt 87 lt 211 gt 107 lt 212 gt PRT lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence 7 10VL_A43V mutant lt 400 gt 87 Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Thr Cys Arg Ala Ser Gln Gly Ser Ser Ala 20 25 30 10 20 30 40 109 WO2007 108559 1 2007 9 27 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Val Pro Lys Leu Leu 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ser Ser Leu Pro 65 70 75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Leu 85 90 95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105 lt 210 gt 88 lt 211 gt 107 lt 212 gt PRT lt 213 gt Artificial Sequence lt 220 gt 223 Description of Artificial Sequence 7 10VL_Y94F mutant lt 400 gt 88 Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Thr Cys Arg Ala Ser Gln Gly Ser Ser Ala 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu
221. evant passages Relevant to claim No JP 2002 512776 A SmithKline Beecham 1 12 2002 08 05 02 1999 055369 A1 amp EP 1073464 A1 2002 0062009 A1 amp EP 1073464 B1 69920897 E amp ES 2230848 T3 6936698 B2 amp US 2005 0208625 1 69920897 T2 WO 1999 010494 A2 GENENTECH INC 04 March 1999 04 03 99 amp AU 1998 088312 A amp EP 1009831 A2 amp JP 2001 513999 A amp US 6342220 B1 amp AU 755822 B amp AU 2003 201322 1 amp AU 2003 201322 B2 Form PCT ISA 210 continuation of second sheet April 2005 177 02007 108559 1 2007 9 27 INTERNATIONAL SEARCH REPORT International application No PCT JP2007 056536 Continuation of B FIELDS SEARCHED Electronic data base consulted during the international search name of data base and where practicable search terms used SwissProt PIR Geneseg Form PCT ISA 210 extra sheet April 2005 178 WO2007 108559 1 2007 9 27 INTERNATIONAL SEARCH REPORT International application No PCT JP2007 056536 Box No II Observations where certain claims were found unsearchable Continuation of item 2 of first sheet This international search report has not been established in respect of certain claims under Article 17 2 a for the following reasons 1 Claims Nos because they relate to subject matter not required to be searched by this Authority namely 2 0 Claims Nos because they relate to parts of the int
222. fee this Authority did not invite payment of any additional fee 3 0 As only some of the required additional search fees were timely paid by the applicant this international search report covers only those claims for which fees were paid specifically claims Nos No required additional search fees were timely paid by the applicant Consequently this international search report is restricted to the invention first mentioned in the claims it is covered by claims Nos 1 12 Remark on Protest 0 The additional search fees were accompanied by the applicant s protest and where applicable the payment of a protest fee 0 The additional search fees were accompanied by the applicant s protest but the applicable protest fee was not paid within the time limit specified in the invitation Form PCT ISA 210 continuation of first sheet 2 April 2005 179 02007 108559 Al 2007 9 27 INTERNATIONAL SEARCH REPORT International application No PCT JP2007 056536 Continuation of Box No III of continuation of first sheet 2 Rule 13 3 Now examinationis made about the common feature among the inventions of claims 1 13 14 15 17 18 and 19 which are independent claims The inventions of claims 14 15 17 and 18 relate to specific DNA comprising anucleotide sequence encoding specific amino acid sequence selected from the group consisting of the amino acid sequence depicted in a specific SEQ ID number and an amino acid s
223. g 180 aaaccaggga aagttcctaa gctcctgatc tatgatgcct ccagtttgga aagtggggtc 240 ccatcaaggt tcagcggcag tggatctggg acagatttca ctctcaccat cagcagcctg 300 cagcctgaag attttgcaac ttattactgt caacagttta atagttaccc gtggacgttc 360 ggccaaggga ccaaggtgga aatcaaacgt 390 lt 210 gt 34 lt 211 gt 130 lt 212 gt PRT lt 213 gt Homo sapiens lt 400 gt 34 Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp 1 5 10 15 Leu Pro Gly Ala Arg Cys Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser 20 25 30 Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45 Gln Gly Ile Ser Ser Ala Leu Ala Trp Tyr Gln Gin Lys Pro Gly Lys 50 55 60 Val Pro Lys Leu Leu Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val 65 70 15 80 Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 85 90 95 Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln 100 105 110 Phe Asn Ser Tyr Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu 115 120 125 Lys Arg 130 lt 210 gt 35 10 20 30 40 lt 211 gt 429 lt 212 gt DNA lt 213 gt Homo lt 400 gt 35 atggagttgg gtgcaactgg tgtgcagcct gactctgtga Caaatgaaca ttcggggagt gtctcctca lt 210 gt 36 lt 211 gt 143 lt 212 gt PRT lt 213 gt Homo lt 400 gt 36 sapiens gactgagctg tggagtgigg ctggattcac tggagtgggt agggccgatt gtctgagagc EggEggaaacta sapiens gattttcctt gggaggettg ctttgatgat ctcaggtatt Caccatctcc tgaggacacg ctacggtatg
224. gagtgggt ctcaggtatt agttggaaca gtggtagcat aggctatgcg 240 gactctgtga agggccgatt caccgtttcc agagacaacg Ccaagaactc cctgtatctg 300 caaatgaaca gtctgagagc tgaggacacg gccttatatt actgtgcaaa agccctatgg 360 ttcggggagt tcccccacta ctacggtatg gacgtctggg gccaagggac Cacggtcacc 420 gtctcctca 429 lt 210 gt 28 lt 211 gt 143 lt 212 gt lt 213 gt Homo sapiens lt 400 gt 28 Met Glu Leu Gly Leu Ser Trp Ile Phe Leu Val Ala Ile Leu Lys Gly 1 5 10 15 Val Gln Cys Glu Glu Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln 20 25 30 Pro Gly Arg Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe 35 40 45 10 20 30 40 71 02007 108559 1 2007 9 27 Asp Asp Tyr Ala Met Tyr Trp Val Arg Gln Val Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ala Lys Asn 85 90 95 Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu 100 105 110 Tyr Tyr Cys Ala Lys Ala Leu Trp Phe Gly Glu Phe Pro His Tyr Tyr 115 120 125 Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 130 lt 210 gt 29 lt 211 gt 390 lt 212 gt DNA lt 213 gt Homo lt 400 gt 29 atggacatga agatgtgcca gtcaccatca aaaccaggga ccatcaaggt cagcctgaag lt 210 gt 30 lt 211 gt 130 lt 212 gt lt 213 gt lt 400 gt 30 sapiens gggtccccgc tccagttgac cttgccgggc aagctcctaa tcagcggcag
225. gcc lt 210 gt 41 lt 211 gt 45 212 DNA lt 213 gt Artificial Sequence Ile Lys 45 40 10 20 30 40 78 lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 41 agagagagag gtcgaccacc atggagttgg gactgagctg gattt lt 210 gt 42 lt 211 gt 37 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 42 agagagagag gctagctgag gagacggtga ccgtggt lt 210 gt 43 lt 211 gt 42 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 43 agagagagag atctctcacc atggacatga gggtccccgc tc lt 210 gt 44 lt 211 gt 40 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 44 agagagagag cgtacgtttg atctccacct tggtccctcc WO2007 108559 Al 2007 9 27 45 10 47 20 30 42 40 40 79 lt 210 gt 45 lt 211 gt 41 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 45 agagagagag atctctcacc atgagggtcc ccgctcagct c lt 210 gt 46 211 gt 42 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 46 agagagagag cgtacgtttg atttccacct tggtcccttg ge lt 210 gt 47 lt 211 gt 45
226. gg cgccctgacc agcggcgtgc acaccttccc ggctgtccta cagtcctcag 180 gactctactc acacctgcaa aaaccccact acacacctcc gcccacggtg cacctgaact ttatgatttc ccgaggtcca tgcgggagga aggactggct ccatcgagaa tgcccccatc gcttctaccc acaacaccac ccgtggacaa ctctgcacaa lt 210 gt 57 lt 211 gt 27 lt 212 gt DNA cctcagcagc cgtgaatcac tggtgacaca cccgtgccca cccagagccc cctgggagga ccggacecct gttcaagtgg gcagtacaac gaacggcaag aaccatctcc ccgggaggag cagcgacatc gcctcccatg gagcaggtgg ccgctacacg gtggtgaccg aagcccagca actcacacat cggtgcccag aaatcttgtg ccgtcagtct gaggtcacgt tacgtggacg agcacgttcc gagtacaagt aaaaccaaag atgaccaaga gccgtggagt ctggactccg cagcagggga 213 gt Artificial Sequence lt 220 gt 84 tgccctccag acaccaaggt Ecccacggtg agcccaaatc acacacctcc tectettece gcgtggtggt gcgtggaggt gtgtggtcag gcaaggtctc gacagecccg accaggtcag gggagagcaa acggctcctt acatcttctc tctccctgtc WO2007 108559 Al 2007 9 27 cagtttgggc ggacaagaga ttgtgacaca ggacgtgagc gcataatgcc cgtcctcacc caacaaagcc agaaccacag cctgacctgc tgggcagccg cttcctctac atgctccgtg tccgggtaaa lt 223 gt Description of Artificial Sequence primer lt 400 gt 57 gggtacgtcc tcacattcag tgatcag lt 210 gt 58 lt 211 gt 30 212 DNA 213 Artificial Sequence 220 223 Description of Artificial Sequence primer 400 58 gtcttcgtgg
227. gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 53 agagagagag gtcgaccacc atggagttgg gactgagctg gattt lt 210 gt 54 lt 211 gt 38 lt 212 gt DNA lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence primer lt 400 gt 54 agagagagag gctagctgag gagacggtga ccgtggtc 210 gt 55 lt 211 gt 376 212 gt PRT lt 213 gt Homo sapiens WO2007 108559 Al 2007 9 27 10 37 20 45 30 38 40 lt 400 gt 55 Ser Thr Lys Gly Pro Ser Val Phe Pro 1 5 Thr Ser Gly Gly Thr Ala Ala Leu Gly 20 25 Pro Glu Pro Val Thr Val Ser Trp Asn 35 40 Val His Thr Phe Pro Ala Val Leu Gln 50 55 Ser Ser Val Val Thr Val Pro Ser Ser 65 70 Thr Cys Asn Val Asn His Lys Pro Ser 85 Val Glu Leu Lys Thr Pro Leu Gly Asp 100 105 82 WO2007 108559 Al 2007 9 27 Leu Ala Pro Cys Ser 10 Cys Leu Val Lys Asp 30 Ser Gly Ala Leu Thr 45 Ser Ser Gly Leu Tyr 60 Ser Leu Gly Thr Gln 75 Asn Thr Lys Val Asp 90 Thr Thr His Thr Cys 110 Arg Ser 15 Tyr Phe Ser Gly Ser Leu Thr Tyr 80 Lys Arg 95 Pro Arg Cys Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg Cys 115 120 Pro Glu Pro Lys Ser Cys Asp Thr Pro 130 135 Glu Pro Lys Ser Cys Asp Thr Pro Pro 145 150 Pro Glu Leu Leu Gly Gly Pro Ser Val 165 Lys Asp Thr Leu Met Ile Ser Arg Thr 180 185 Val Asp Val Ser His Glu Asp Pro Glu 195
228. le Ser Arg Asp Asn Ala Lys Asn 85 90 95 Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu 100 105 110 Tyr Tyr Cys Ala Lys Asn Leu Trp Phe Gly Glu Phe Arg Tyr Trp Tyr 115 120 125 Phe Asp Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser Ala Ser 130 135 140 Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr 145 150 155 160 Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro 165 170 175 Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val 180 185 190 His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser 195 200 205 Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr 210 215 220 Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val 225 230 235 240 Glu Ser Lys Thr Pro Leu Gly Asp Thr Thr His Thr Cys Pro Pro Cys 245 250 255 Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 260 265 270 10 20 30 40 104 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 275 280 Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val 290 295 300 Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 305 310 315 Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val 325 330 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 340 345 Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser 355 360 Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 370 375 380 Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 385 390 395 Pro Ser Asp Ile Ala Val Glu Trp Glu Se
229. r Ala His Lys Asp Pro Asn Ala Ile Phe 115 120 125 Leu Ser Phe Gin His Leu Leu Arg Gly Lys Val Arg Phe Leu Met Leu 130 135 140 Val Gly Gly Ser Thr Leu Cys Val Arg Arg Ala Pro Pro Thr Thr Ala 145 150 155 160 Val Pro Ser lt 210 gt 2 lt 211 gt 123 lt 212 gt lt 213 gt Homo sapiens lt 400 gt 2 Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Lys Asn Leu Trp Phe Gly Glu Phe Arg Tyr Trp Tyr Phe Asp Leu 100 105 110 10 20 30 40 57 WO2007 108559 Al 2007 9 27 Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser 115 120 lt 210 gt 3 lt 211 gt 107 lt 212 gt PRT lt 213 gt Homo sapiens lt 400 gt 3 Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Glu Asp Phe Ala
230. r Asn Gly 405 410 Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 420 425 Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp 435 440 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 450 455 460 Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys 465 470 lt 210 gt 83 lt 211 gt 711 WO2007 108559 1 2007 9 27 Glu Val Thr Cys 285 Gln Phe Asn Trp Lys Pro Arg Glu 320 Leu Thr Val Leu 335 Lys Val Ser Asn 350 Lys Ala Lys Gly 365 Ser Gln Glu Glu Lys Gly Phe Tyr 400 Gln Pro Glu Asn 415 Gly Ser Phe Phe 430 Gln Glu Gly Asn 445 Asn His Tyr Thr 10 20 30 40 lt 212 gt 213 gt Artificial Sequence lt 220 gt 105 WO2007 108559 Al 2007 9 27 lt 223 gt Description of Artificial Sequence 7 1064344uhm L chain lt 400 gt 83 atggacatga agatgtgcca gtcaccatca aaaccaggga ccatcaaggt cagcctgaag CCgccatctg ttctatccca tcccaggaga ctgacgctga cagggcctga 210 84 lt 211 gt 236 lt 212 gt gggtccccgc tccagttgac cttgccgggc aagctcctaa tcagcggcag attttgcaac ccaaggtgga atgagcagtt gagaggccaa gtgtcacaga gcaaagcaga gctcgcccgt tcagctcctg ccagtctcca aagtcagggc gctcctgatc tggatctggg ttattactgt gatcaaacgt gaaatctgga agtacagtgg gcaggacage ctacgagaaa cacaaagagc lt 213 gt Artificial Sequence 220 gggCttctgc tcctccctgt attagcagtg tatgatgcct acagatttca caacagttta acggtggctg actgcctctg aaggtggata aaggacagca
231. re the inventions of claims 1 13 14 15 17 18 and 19 do not comply with the requirement of unity of invention According to the decision stated above it is obvious that a group of inventions which are claimed as alternatives in the inventions of Claims 14 15 17 and 18 cannot also be regarded as a group of inventions linked to forma single general inventive concept considered that claims 14 15 17 and 18 include different four four eight and eight DNA related inventions respectively In conclusion the present application includes the following 27 inventions in total 1 an invention relating to claim 1 and parts of claims 2 12 referring to claim 1 ii an invention relating to claim 13 iii four inventions relating to claim 14 which include four groups of inventions four inventions relating to claim 15 and part of claim 16 referring to claim 15 which include four groups of inventions eight inventions relating to claim 17 which include eight groups of inventions vi eight inventions relating to claim 18 which include eight groups of inventions and vii an invention relating to claim 19 and a part of claim 20 referring to claim 19 Form PCT ISA 210 extra sheet April 2005 180 02007 108559 1 2007 9 27 PCT JP2007 056536 I PC C12N15 09 2006
232. rg Trp Gln Glu Gly Asn Val Phe Ser 290 295 300 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 305 310 315 320 39 Leu Ser Leu Ser Leu Gly Lys 325 lt 210 gt 81 lt 211 gt 1425 lt 212 gt DNA lt 213 gt Artificial Sequence 40 220 lt 223 gt Description of Artificial Sequence 7 1064344uhm H chain lt 400 gt 81 atggagttgg gactgagctg gattttcctt ttggctattt taaaaggtgt ccagtgtgaa 60 gtgcagctgg tggagtctgg gggaggcttg gtacagcctg gcaggtccct gagactctcc 120 tgtgcagcct gactctgtga Caaatgaaca ttcggggagt tcctcagcta tccgagagca gtgtcgtgga tcctcaggac aagacctaca gagtccaaaa ttcgaggggg tcccggaccc cagttcaact gagcagttca ctgaacggca aaaaccatct tcccaggagg cccagcgaca acgcctcccg aagagcaggt aaccactaca lt 210 gt 82 lt 211 gt 474 lt 212 gt PRT ctggattcac tggagtgggt agggccgatt gtctgagagc tccgttactg gcaccaaggg cagccgccct actcaggcgc tctactccct cctgcaacgt ccccacttgg gaccatcagt ctgaggtcac ggtacgtgga acagcacgta aggagtacaa ccaaagccaa agatgaccaa tcgccgtgga tgctggactc cacagaagag ctttgatgat ctcaggtatt caccatctcc tgaggacacg gtacttcgat gccatcggtc gggctgcctg cctgaccagc cagcagcgtg agatcacaag tgacacaact cttcctgttc gtgcgtggtg tggcgtggag ccgtgtggtc gtgcaaggtc agggcagccc gaaccaggtc gtgggagagc cgacggctcc gaatgtcttc cctctccctg lt 213 gt Artificial Sequence lt 220 gt 102 tatgccatge agttggaata agagacaacg gccttgtatt ctctggggcc
233. s Thr Lys 180 185 190 Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu 200 205 Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 215 220 Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys 230 235 Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 245 250 Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 260 265 270 Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 280 285 Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 295 300 Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 310 315 Ser Cys Ser Val Met His Glu Ala Leu His Asn 325 330 Ser Leu Ser Leu Ser Leu Gly Lys 340 345 lt 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence G4344uh Pro Arg Thr Val Val Ser Ala Lys 240 Gln Glu 255 Gly Phe Pro Glu Ser Phe Glu Gly 320 His Tyr 335 10 20 30 40 97 WO2007 108559 Al 2007 9 27 lt 400 gt 78 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser 1 Thr 1 Ala Leu 1 Leu Val Lys Asp 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Leu Met Ile Ser 50 55 60 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp 65 70 75 80 Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 85 90 95 Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val 100 105 110 Val Ser Val Leu Thr Val Leu His Gln A
234. sp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val Glu Leu Lys Thr Pro Leu Gly Asp Thr Thr His Thr Cys Pro 100 105 110 Arg Cys Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg 115 120 125 Cys Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg Cys 130 135 140 Pro Glu Pro Lys Ser Cys Asp Thr Pro Pro Pro Cys Pro Arg Cys Pro 145 150 155 160 Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 94 WO2007 108559 Al 2007 9 27 165 170 175 Pro Lys Asp Thr Leu Met Ser Arg Thr Pro Glu Val Thr Cys Val 180 185 190 Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr 195 200 205 Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 210 215 220 Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 225 230 235 240 Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 245 250 255 Gly Leu Pro Ser Ser Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln 260 265 270 Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met 275 280 285 Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro 290 295 300 Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn 305 310 315 320 Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 325 330 335 Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val 340 345 350 Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln 355 360 365 Lys Ser Leu Ser Leu Ser
235. sp Trp Leu Asn Gly Lys Glu 115 120 125 Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys 130 135 140 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 145 150 155 160 Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 165 170 175 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 180 185 190 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 195 200 205 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 210 215 220 98 WO2007 108559 Al 2007 9 27 Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu 225 230 235 240 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly 245 250 255 Lys lt 210 gt 79 lt 211 gt 332 lt 212 gt PRT 213 gt Artificial Sequence lt 220 gt lt 223 gt Description of Artificial Sequence G4344uhm lt 400 gt 79 Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg 1 5 10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45 Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60 Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 65 70 75 80 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95 Arg Val Glu Ser Lys Thr Pro Leu Gly Asp Thr Thr His Thr Cys Pro 100 105 110 Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly
236. tgcatctgt aggagacaga 120 gtcaccatca cttgccgggc aagtcagggc attagcagtg ctttagcctg gtatcagcag 180 aaaccaggga aagctcctaa gctcctgatc tatgatgcct ccagtttgga aagtggggtc 240 ccatcaaggt tcagcggcag tggatctggg acagatttca ctctcaccat cagcagcctg 300 cagcctgaag attttgcaac ttattactgt caacagttta atagttaccc gctcactttc 360 ggcggaggga ccaaggtgga gatcaaa 387 210 26 lt 211 gt 142 lt 212 gt lt 213 gt Homo sapiens 400 26 Met Glu Leu Gly Leu Ser Trp Ile Phe Leu Leu Ala Ile Leu Lys Gly 1 5 10 15 Val Gln Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln 20 25 30 Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 Asp Asp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn 85 90 95 70 02007 108559 Al 2007 9 27 Ser Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu 100 105 110 Tyr Tyr Cys Ala Lys Asn Leu Trp Phe Gly Glu Phe Arg Tyr Trp Tyr 115 120 125 Phe Asp Leu Trp Gly Arg Gly Thr Leu Val Thr Val Ser Ser 130 135 140 lt 210 gt 27 lt 211 gt 429 lt 212 gt DNA lt 213 gt Homo sapiens lt 400 gt 27 atggagttgg gactgagctg gattttcctt gtggctattt taaaaggtgt ccagtgtgaa 60 gagcagctgg tggagtctgg gggaggcttg gtacagcctg gcaggtccct gagactctcc 120 tgtacagcct ctggattcac ctttgatgat tatgccatgt actgggtccg gcaagttcca 180 gggaagggcc tg
237. ttccccctgg gtcaaggact ggcgtgcaca gtgaccgtgc cccagcaaca cacacatgcc gtggacgtga gtgcataatg agcgtcctca tccaacaaag cgagagccac agcctgacct aatgggcage ttcttcctct tcatgctccg tctctgggta actgggtccg gtggtagcat ccaagaactc actgtgcaaa gtggcaccct cgccctgctc acttccccga ccttcccggc cctccagcag ccaaggtgga caccatgccc ccaaggacac gccaggaaga ccaagacaaa Ccgtcctgca gCctcccgtc aggtgtacac gcctggtcaa cggagaacaa acagcaggct tgatgcatga aatga WO2007 108559 1 2007 9 27 gcaagctcca aggctatgcg cctgtatctg aaatctatgg ggtcactgtc caggagcacc accggtgacg tgtcctacag cttgggcacg caagagagtt agcacctgag tctcatgatc ccccgaggtc gCCgCgggdg CCaggactgg ctccatcgag cctgccccca aggcttctac ctacaagacc aaccgtggac ggCtctgcac lt 223 gt Description of Artificial Sequence 7 1064344uhm H chain lt 400 gt 82 Met Glu Leu Gly Leu Ser Trp Ile Phe Leu Leu Ala Ile Leu Lys Gly 1 5 10 15 Val Gln Cys Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln 30 20 25 Pro Gly Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 35 40 45 180 240 300 360 420 480 540 600 660 120 180 840 900 960 1020 1080 1140 1200 1260 1320 1380 1425 10 20 30 40 103 02007 108559 1 2007 9 27 Asp Asp Tyr Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 50 55 60 Glu Trp Val Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala 65 70 75 80 Asp Ser Val Lys Gly Arg Phe Thr I
238. uoz G 8 06 9 1 A 1 9 1 8 S S d 1 A AS S 1 S 419 S S b 1 4851 991 214 301 922 001 DIL 998 999 283 961 280 SIL 901 996 918 199 LOY 998 319 LUO 228 9bH 209 911 398 DIL 222 919 228 919 919 29H 29H 212 201 281 212 YOO HIL 221 983 HLI 319 209 V d d 1 9 S 1 31 9 9 S N S A 1 A d 3 d d A 12858955 493 208 991 919 292 922 921 001 999 692 10H OLI 22H 208 280 904 383 209 112 909 918 812 199 995 311 208 383 919 209 208 338 ALI 339 208 991 931 919 938 918 999 BHO 333 ILA 281 225 A 19 9 7 v wv L 8 3 S8 4 S 3 d WV 31 d 4 A S 311 954 933 993 999 LOL SIL 212 001 931 321 55H 999 999 999 BBH 2808 199 219 919 DOL 999 OLI 229 200 YOY DOH 990 221 JOY 29H 99H 221 201 229 999 813 200 211 919 991 822 ZG 9 4 1 S S S A 1 A 1 O0 N 9 A 4d A A 3 94521 222 211 901 994 499 108 9H3 983 OHD 901 933 829 999 22H ULI SHY OLY JOU 91H 212 LT 999 298 m BJL 221 219 12 219 612 208 299 190 399 991 915 189 ILL 281 991 381 193 211 980 24 9 1 N a y 3 1 v 1 3 1 5 N W 1 1 435 222 OB LUO HLL LLL 193 OLH HLY 2HH 002 291 OLI 212 HOO LIL HIL 911 JUL LIO 900 211 901 BLI lHH 599 191 911 229 996 2H9 120 910 LOY 2BH 918 883 013 812 ZAE S N A v N
239. y 1 5 Ser Leu Arg Leu Ser Cys Ala Ala 20 Ala Met His Trp Val Arg Gln Ala 35 40 Ser Gly Ile Ser Trp Asn Ser Gly 50 55 25 30 Gly Lys Val Pro Lys Leu Leu 45 Gly Val Pro Ser Arg Phe Ser Gly 60 Leu Thr Ile Ser Ser Leu Gln Pro 75 80 Gln Gln Phe Asn Ser Tyr Pro Trp 90 95 Glu Ile Lys Arg 105 Gly Gly Leu Val Gln Pro Gly Arg 10 15 Ser Gly Phe Thr Phe Asp Asp Tyr 25 30 Pro Gly Lys Gly Leu Glu Trp Val 45 Ser Ile Gly Tyr Ala Asp Ser Val 60 10 20 30 40 61 02007 108559 Al 2007 9 27 Lys Gly Arg Phe Thr Ser Arg Asp Asn Ala Lys Asn Leu 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Lys Pro Ile Trp Phe Gly Glu Trp Gly Asn Tyr Tyr Gly Met Asp 100 105 110 Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 lt 210 gt 9 lt 211 gt 109 lt 212 gt PRT lt 213 gt Homo sapiens lt 400 gt 9 Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Asp Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95 Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg 100 105 10 20

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