ederation of American Societies for Experimental Biology 10,1227-1232 (1996).
[0209] 18. Mendez? M. J. et al. Functional transplant of megabase human immunoglobulin loci recapitulates human antibody response in mice. Nature genetics 15,146-156(1997) ·
[0210] 19.Davies? N. P. et al.Creation of mice expressing human antibody light chains by introduction of a yeast artificial chromosome containing the core region of the human immunoglobulin kappa locus.Biotechnology(N Y) 11， 911-914(1993).
[0211] 20. Davies，N. P.，Popov，A. V.，Zou，X. &Briiggemann，M. Human antibody repertoires in transgenic mice Manipulation and transfer of YACs·· IRL Oxford， 59-76(1996).
[0212] 21. Lonberg，N. et al. Antigen-specific human antibodies from mice comprising four distinct genetic modifications. Nature 368,856-859 (1994) ·
[0213] 22. Nicholson? I. C. et al. Antibody repertoires of four-and five-feature translocus mice carrying human hmmunoglobulin heavy chain and kappa and lambda light chain yeast artificial chromosomes. J Immunol 163,6898-6906(1999) ·
[0214] 23. Pruzina? S. et al. Human monoclonal antibodies to HIV-I gpl40 from mice bearing YAC-based human immunoglobulin transloci. Protein engineering， design&selection :PEDS 24,791-799(2011).
[0215] 24. Wagner，S. D.，Gross，G.，Cook，G. P.，Davies，S. L &Neuberger，M. S. Antibody expression from the core region of the human IgH locus reconstructed in transgenic mice using bacteriophage Pl clones. Genomics 35,405-414 (1996) ·
[0216] 25. Popov，A. V.，Butzler，C.，F(xiàn)rippiat，J. P.，Lefranc，Μ. P. &Bruggemann， M.Assembly and extension of yeast artificial chromosomes to build up a large locus.Gene 177,195-201 (1996).
[0217] 26. Mundt? C. A. et al. Novel control motif cluster in the IgH delta-gamma 3 interval exhibits B cell-specific enhancer function in early development. J Immunol 166,3315-3323 (2001).
[0218] 27. Lefranc，M. -P. &Lefranc，G. The immunoglobulin factsbook. FactsBook Series，Academic Press，GB，45-68(2001).
[0219] 28. Reynaud? S. et al. Interallelic class switch recombination contributes significantly to class switching in mouse B cells. J Immunol 174， 6176-6183(2005).
[0220] 29. Bruggemann，M.，Teale，C.，Clark，M.，Bindon，C. &Waldmann，H. A matched set of rat/mouse chimeric antibodies. Identification and biological properties of rat H chain constant repons mu，gamma 1，gamma 2a，gamma 2b，gamma 2c，epsilon，and alpha. J Immunol 142,3145-3150 (1989).
[0221] 30. Bazin，H. ,Beckers，A. &Querinjean，P. Three classes and four (sub)classes of rat immunoglobulins :IgM，IgA，IgE and IgGl，IgG2a，IgG2b，IgG2c. European journal of immunology 4? 44-48 (1974).
[0222] 31. McGhee，J. R.，Michalek，S. M. &Ghanta，V. K. Rat immunoglobulins in serum and secretions purification of rat IgM，IgA and IgG and their quantitation in serum，colostrum，milk and saliva. Immunochemistry 12,817-823(1975).
[0223] 32. Shansab，M.，Eccleston，J. M. &Selsing，E. Translocation of an antibody transgene requires AID and occurs by interchromosomal switching to all switch regions except the mu switch region.European journal of immunology 41， 1456-1464(2011).
[0224] 33. Bruggemann，M. et al. Human antibody production in transgenic mice ： expression from 100 kb of the human IgH locus. European journal of immunology 21，1323-1326(1991).
[0225] 34. Vincent-Fabert，C. et al. Genomic deletion of the whole IgH 3 regulatory region(hs3a，hsl，2，hs3b，and hs4)dramatically affects class switch recombination and Ig secretion to all isotypes. Blood 116,1895-1898 (2010) ·
[0226] 35. Davis? C. G. ? Gallo? M. L. &Corvalan? J. R. Transgenic mice as a source of fully human antibodies for the treatment of cancer. Cancer metastasis reviews 18,421-425 (1999).
[0227] 36. Lonberg? N. Fully human antibodies from transgenic mouse and phage display platforms. Current opinion in immunology 20,450-459 (2008)·
[0228] 37.Chen?K. C. ?A. New Insights into the Enigma of Immunoglobulin D. Immunol Rev 237,160-179(2010).
[0229] 38. Popov，A. V.，Zou，X.，Xian，J.，Nicholson，I. C. &Bruggemann，M. A human immunoglobulin lambda locus is similarly well expressed in mice and humans. The Journal of experimental medicine 189,1611-1620 (1999) ·
[0230] 39. Marchuk，D. &Collins，F(xiàn). S. pYAC-RC，a yeast artificial chromosome vector for cloning DNA cut with infrequently cutting restriction endonucleases. Nucleic acids research 16?7743 (1988).
[0231] 40. Kaur? R. ? Ma? B. &Cormack? B. P. A family of glycosylphosphatidylinosit ol-linked aspartyl proteases is required for virulence of Candida glabrata. Proceedings of the National Academy of Sciences of the United States of America 104,7628-7633 (2007).
[0232] 41. Nelson?D. L. &Brownstein?B. H. YAC libraries ：A users guide. . Freeman and Company，NY (1994).
[0233] 42.Gietz? D. &Woods? R.A. Transformation of yeast by the lithium acetate single-stranded carrier DNA/PEG method. Methods in Microbiology 26,53-66 (1998) ·
[0234] 43.Peterson? K. R. Preparation of intact yeast artificial chromosome DNA for transgenesis of mice. Nature protocols 2?3009-3015 (2007).
[0235] 44. Beverly? S. M. Characterization of the ^unusuali mobility of large circular DNAs in pulsed field-gradient electrophoresis Nucleic acids research 16,925-939 (1988).
[0236] 45. Kawasaki? K. et al. Evolutionary dynamics of the human immunoglobulin kappa locus and the germline repertoire of the Vkappa genes. European journal of immunology 31，1017-1028(2001).
[0237] 46. Sambrook?J. &RusselI?D. W. Molecular Cloning. A laboratory Manual. . Cold Spring Harbor Laboratory Press，NY(2001).
[0238] 47. Gu，H.，Wilson，D. &Inselburg，J. Recovery of DNA from agarose gels using a modified Elutrap. Journal of biochemical and biophysical methods 24， 45-50(1992).
[0239] 48. Yang，X. I，Model，P. &Heintz，N. Homologous recombination based modification in Escherichia coli and germline transmission in transgenic mice of a bacterial artificial chromosome. Nature biotechnology 15,859-865 (1997) ·
[0240] 49. Meisner，L M.，and J. Johnson Protocols for cytogenetic studies of human embryonic stem cells. . Methods 45,133-141 (2008) ·
[0241] 50. Kishiro, Y.，Kagawa，M.，Naito，I. &Sado, Y. A novel method of preparing rat-monoclonal antibody-producing hybridomas by using rat medial iliac lymph node cells.Cell structure and function 20,151-156 (1995) ·
【主權(quán)項(xiàng)】
1. 一種嵌合多核苷酸，其包含至少一個(gè)人免疫球蛋白（Ig)連接（J)區(qū)基因、Ig恒定區(qū) 基因和大鼠3'增強(qiáng)子。2. 如權(quán)利要求1所述的嵌合多核苷酸，其中所述大鼠3'增強(qiáng)子包括如SEQ ID NO: 1所 示的序列。3. 如前述權(quán)利要求中任一項(xiàng)所述的嵌合多核苷酸，其還包含至少一個(gè)人Ig可變（V)區(qū) 基因和/或人Ig多樣性（D)區(qū)基因。4. 如前述權(quán)利要求中任一項(xiàng)所述的嵌合多核苷酸，其中所述恒定區(qū)基因選自由人恒定 區(qū)基因和大鼠恒定區(qū)基因組成的組。5. 如前述權(quán)利要求中任一項(xiàng)所述的嵌合多核苷酸，其中所述恒定區(qū)基因包括大鼠恒定 區(qū)基因。6. 如前述權(quán)利要求中任一項(xiàng)所述的嵌合多核苷酸，其中所述恒定區(qū)基因包括選自由 Cy和Cy組成的組的恒定區(qū)基因。7. 如前述權(quán)利要求中任一項(xiàng)所述的嵌合多核苷酸，其包含與細(xì)菌人工染色體（BAC) Annabel或其部分基本同源的核酸序列。8. 如權(quán)利要求3-7中任一項(xiàng)所述的嵌合多核苷酸，其中所述人Ig V區(qū)包含至少一個(gè)可 從BAC6-VH3-11和/或BAC3分離的人V區(qū)基因。9. 如前述權(quán)利要求中任一項(xiàng)所述的嵌合多核苷酸，其以5'至3'的順序包含核酸序列 (a)和（b): (a) 人Ig可變區(qū)，其包含可從BAC6-VH3-11和/或BAC3分離的呈天然構(gòu)型的人V區(qū)基 因；和 (b) 人Ig連接區(qū)，其包含可從所述細(xì)菌人工染色體（BAC)Annabel分離的呈天然構(gòu)型的 人J區(qū)基因。10. 如權(quán)利要求3-9中任一項(xiàng)所述的嵌合多核苷酸，其中所述人免疫球蛋白可變區(qū)、所 述人免疫球蛋白多樣性區(qū)、所述人免疫球蛋白連接區(qū)、所述免疫球蛋白恒定區(qū)和所述大鼠 3'增強(qiáng)子中的每一個(gè)處于圖Ia中所示的相對(duì)位置。11. 如權(quán)利要求10所述的嵌合多核苷酸，其包含與如SEQ ID NO: 2所示的核酸序列基 本同源的核酸序列。12. 如權(quán)利要求10所述的嵌合多核苷酸，其包含與如SEQ ID NO: 11所示的核酸序列基 本同源的核酸序列。13. 如權(quán)利要求3-10中任一項(xiàng)所述的嵌合多核苷酸，其中所述V-D-J區(qū)重排并形成編 碼重鏈可變結(jié)構(gòu)域的完整外顯子。14. 一種嚙齒動(dòng)物細(xì)胞，其包含如前述權(quán)利要求中任一項(xiàng)所述的嵌合多核苷酸。15. 如權(quán)利要求13所述的嚙齒動(dòng)物細(xì)胞，其還包含編碼功能性免疫球蛋白的多核苷 酸，所述多核苷酸包含V-、J-和恒定區(qū)基因以及與選自由SEQ ID NO: 6和SEQ ID NO: 7組 成的組的核酸序列基本同源的核酸序列。
【專(zhuān)利摘要】本發(fā)明涉及可用于在嚙齒動(dòng)物中優(yōu)化產(chǎn)生具有人獨(dú)特型的功能性免疫球蛋白的多核苷酸，尤其是嵌合多核苷酸。本發(fā)明還涉及包含此類(lèi)多核苷酸的嚙齒動(dòng)物。
【IPC分類(lèi)】C12N15/85, A01K67/027, C07K16/00
【公開(kāi)號(hào)】CN104994729
【申請(qǐng)?zhí)枴緾N201380070457
【發(fā)明人】M·布魯格曼, R·布洛夫, M·J·奧斯伯恩, B·馬
【申請(qǐng)人】Omt公司
【公開(kāi)日】2015年10月21日
【申請(qǐng)日】2013年12月13日
【公告號(hào)】CA2895144A1, EP2931030A2, US20150113668, WO2014093908A2, WO2014093908A3