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(12) United States Patent (10) Patent N0.: US 8,283,294 B2 Kastrup Et A] US008283294B2 (12) United States Patent (10) Patent N0.: US 8,283,294 B2 Kastrup et a]. (45) Date of Patent: Oct. 9, 2012 (54) METHOD FOR CLONING COGNATE FOREIGN PATENT DOCUMENTS ANTIBODIES FR 2724393 3/1996 W0 WO 92/15678 Al 9/1992 (75) Inventors: Jesper Kastrup, Stenlose (DK); Lars S. W0 WO 93/03151 Al 2/1993 Nielsen, Niva (DK); Per-Johan Meijer, W0 WO 93/20227 A1 10/1993 Copenhagen (DK) W0 WO 94/08008 Al 4/1994 W0 WO 95/20401 Al 8/1995 (73) Assignee: Symphogen A/S, Lyngby (DK) W0 WO 96/08564 Al 4/1996 W0 WO 98/57994 A2 12/1998 ( * ) Notice: Subject to any disclaimer, the term of this W0 WO 99/16904 Al 4/1999 patent is extended or adjusted under 35 W0 WO 99/29888 Al 5/1999 U.S.C. 154(b) by 122 days. W0 WO 01/89563 A1 11/2001 W0 WO 01/92291 A2 12/2001 (21) Appl.No.: 12/074,066 WO 2004/003019 * 6/2004 (22) Filed: Feb. 29, 2008 W0 WO 2004/061104 A2 7/2004 (65) Prior Publication Data W0 WO 2005/042774 A2 5/2005 W0 WO 2006/007850 Al l/2006 US 2008/0227660 A1 Sep. 18, 2008 W0 WO 2006/007853 A2 l/2006 W0 WO 2007/003041 Al l/2007 W0 WO 2007/101441 Al l/2007 Related US. Application Data W0 WO 2007/065433 A2 6/2007 (60) Provisional application No. 60/904,772, ?led on Mar. OTHER PUBLICATIONS 5, 2007. Rudikoff et al (Proc Natl Acad Sci USA 1982 vol. 79 p. 1979).* Ward et al. (Nature 341:544-546 (1989)).* (30) Foreign Application Priority Data Smith-Gill et al. (J. Immunol. 139:4135-4144 (1987)).* Kumar et al. (J. Biol. Chem. 275:35129-35136 (2000)).* Mar. 1, 2007 (DK) ............................... .. 2007 00316 Song et al. (Biochem Biophys Res Comm 268:390-394 (2000)).* Soderlind et al. (Gene 160:269-272 (1995).* (51) Int. Cl. Soderlind et al. (Immunotechnology 4:279-285 (Mar. 1999).* Ardavin, C., et al., “B Cell Response After MMTV Infection: C40B 40/08 (2006.01) Extrafollicular Plasmablasts Represent the Main Infected Population C40B 50/06 (2006.01) and Can Transmit Viral Infection,” J'. Immunol. 162:2538-2545, The (52) US. Cl. ........................ .. 506/17; 506/26; 424/133.1 American Association of Immunologists (1999). (58) Field of Classi?cation Search .................. .. 506/17, Babcook, 18., et al., “A novel strategy for generating monoclonal antibodies from single, isolated lymphocytes producing antibodies of 506/26; 424/133.1 de?ned speci?cities,” Proc. Natl. Acad. Sci. USA 93:7843-7848, See application ?le for complete search history. National Academy of Science (1996). Bregenholt, S., et al., “Recombinant Human Polyclonal Antibodies: (56) References Cited A New Class of Therapeutic Antibodies Against Viral Infections,” Curr". Pharm. Des. 12:2007-2015, Bentham Science Publishers (Jan. U.S. PATENT DOCUMENTS 2006). Haurum, J.S., “Recombinant polyclonal antibodies: the next genera 4,683,202 A 7/1987 Mullis 5,698,435 A 12/1997 Robinson et al. tion of antibody therapeutics?,” Drug Discov. Today 11:655-660, 5,882,856 A 3/1999 Shuber Elsevier Science (Jul. 2006). 5,912,172 A 6/1999 Eshhar et a1. Liu, A.Y., et al., “Expression of mousezzhuman immunoglobulin 6,096,878 A 8/2000 Honjo et a1. heavy-chain cDNA in lymphoid cells,” Gene 54:33-40, Elsevier Sci 6,111,166 A 8/2000 van de Winkel ence (1987). 6,258,529 B1 7/2001 BerdoZ et al. Liu, M.-L., et al., “Chronic idiopathic myelo?brosis terminating in 6,538,124 B1 3/2003 Idusogie et al. extramedullary anaplastic plasmacytoma,”Leuk. Lymphoma 47:315 6,849,259 B2 2/2005 Haurum et al. 322, Taylor & Francis (Feb. 2006). 6,867,021 B2 3/2005 Maes et a1. 6,994,963 B1 2/2006 Murphy et a1. (Continued) 7,749,697 B2 7/ 2010 OleksieWicZ et al. Primary Examiner * Lynn Bristol 2001/0027249 A1 10/2001 Le et al. 2002/0102604 A1 8/ 2002 Milne Edwards et a1. (74) Attorney, Agent, or Firm * Sterne, Kessler, 2003/0096343 A1 5/ 2003 Robinson et al. Goldstein & Fox P.L.L.C. 2003/0104477 A1 6/ 2003 Buechler et al. (57) ABSTRACT 2003/0175837 A1 9/2003 Le et al. 2004/0067532 A1 4/2004 Zhu et al. The invention relates to a procedure for linking cognate pairs 2005/0180967 A1 8/ 2005 Haurum et al. ofVH and VL encoding sequences from a population of cells 2006/0228350 A1 10/2006 Wu et al. enriched in particular surface antigen markers. The linking 2006/0275766 A1 12/2006 Haurum et al. procedure involves a multiplex molecular ampli?cation pro 2007/0003515 A1 l/2007 Leonard et a1. cedure capable of linking nucleotide sequences of interest in 2007/0141048 A1 6/2007 OleksieWicZ et al. 2008/0069822 A1 3/2008 Jensen et al. connection With the ampli?cation, in particular polymerase 2008/0131882 A1 6/2008 Rasmussen et al. chain reaction (multiplex PCR). The method is particularly 2008/0226630 A1 9/2008 Lantto et a1. advantageous for the generation of cognate pair libraries as 2009/0004192 A1 l/ 2009 Pedersen et al. Well as combinatorial libraries of variable region encoding 2009/0017017 A1 l/2009 Rasmussen et al. sequences from immunoglobulins. The invention also relates 2009/0111142 A1 4/ 2009 Nielsen et al. to methods for generation of chimeric human/non-human 2009/0136498 A1 5/ 2009 Haurum et al. antibodies and expression libraries generated by such meth 2009/0137003 A1 5/ 2009 Tolstrup et al. ods. 2010/0069262 A1 3/ 2010 Nielsen et al. 2010/0310558 A1 12/ 2010 OleksieWicZ et al. 36 Claims, 8 Drawing Sheets US 8,283,294 B2 Page 2 OTHER PUBLICATIONS Database Biotechno, Accession No. 1999129223369, Abstract for Logtenberg, T., “Antibody cocktails: next-generation Tanabe, R., et al., “Expression of myosin heavy chain isoforms in biopharmaceuticals with improved potency,” Trends Biotechnol. porcine muscles determined by multiplex PCR,” Journal of Food 251390-394, Elsevier Science (Sep. 2007). Science 641222-225, Institute of Food Technologists, US (1999). 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Ther. 61905-912, Ashley Publications (Sep. 2006). Guatelli, J. et al., “Isothermal, in vitro ampli?cation of nucleic acids Weissenhorn, W., et al., “Chimerization of antibodies by isolation of by a multienzyme reaction modeled after retroviral replication,” rearranged genomic variable regions by the polymerase chain reac Proc. Natl. Acad Sci. USA 8711874-1878, The National Academy of tion,” Gene 1061273-277, Elsevier Science (1991). Sciences, US (1990). Wiberg, F.C., et al., “Production of Target-Speci?c Recombinant Harris, E. et al., “Typing of Dengue Viruses in Clinical Specimens Human Polyclonal Antibodies in Mammalian Cells,” Biotechnol. and Mosquitoes by Single-Tube Multiplex Reverse Transcriptase Bioeng. 941396-405, Wiley (Jun. 2006). 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