US010392438B2 (12 ) United States Patent ( 10 ) Patent No. : US 10 ,392 ,438 B2 Bennett et al. ( 45 ) Date of Patent : Aug . 27 , 2019 ( 54 ) BISPECIFIC ANTIBODIES 2002/ 0039995 Al 4 / 2002 Gao 2008 /0269466 Al 10 /2008 Humphreys 2010 /0150914 A1 6 /2010 Wang et al . ( 71 ) Applicant: Pfizer Inc. , New York , NY (US ) 2010 /0256340 A1 10 /2010 Brinkmann 2012 /0009621 A11 /2012 Yamasaki ( 72 ) Inventors : Eric M . Bennett, Arlington , MA (US ) ; 2014 /0200331 A1 * 7 /2014 Corper . .. .. .. .. .. CO7K 16 / 36 Nathan Higginson - Scott, Boston , MA 530 / 387 . 3 (US ) ; Lioudmila Tchistiakova , Stoneham , MA (US ) ; Kimberly A . FOREIGN PATENT DOCUMENTS Marquette , Somerville , MA (US ) ; WO 199852976 11 / 1998 Janet E . Paulsen , Londonderry , NH WO 200034317 6 / 2000 (US ) ; Ruth E . Gimeno , Carmel , IN WO 2009089004 7 / 2009 (US ) wo 20110117653 9 / 2011 WO 2011143545 11 / 2011 ( 73 ) Assignee : PFIZER INC ., New York , NY (US ) WO 20130096291 6 / 2013 wo 2014081955 Al 5 / 2014 ( * ) Notice : Subject to any disclaimer, the term of this WO 2014150973 9 / 2014 patent is extended or adjusted under 35 WO 2015173756 A2 11/ 2015 U . S . C . 154 (b ) by 0 days . OTHER PUBLICATIONS ( 21) Appl . No. : 15 / 309, 879 Davies et al : “ Fab Assembly : An analysis of different Ch1 :CL May 13, 2015 combinations ” , Progress in Immunology VI : Sixth International (22 ) PCT Filed : Congress of Immunology, pp . 145 - 149 , 1986 . ( 86 ) PCT No. : PCT/ IB2015 / 053537 Klein et al: “ Progress in overcoming the chain association issue in bispecific heterodimeric IgG antibodies " , MABS , vol. 4 , No . 6 , pp . $ 371 (c )( 1 ), 653 -663 , 2012 . ( 2 ) Date : Nov . 9 , 2016 Lewis et al : “Generation of bispecific IgG antibodies by structure based design of an orthogonal Fab interface ” , Nature Biotechnol ( 87 ) PCT Pub . No. : W02015 /173756 ogy , vol. 32 , No. 2 , pp . 191 - 198 , 2014 . Lewis et al : “ Supplemental Information : Generation of bispecific PCT Pub. Date : Nov . 19 , 2015 IgG antibodies by structure -based design of an orthogonal Fab interface ” , Nature Biotechnology , vol . 32 , No . 2 , 2014 doi: 10 . 1038 / (65 ) Prior Publication Data nbt. 2797 . US 2018 /0179285 A1 Jun. 28 , 2018 Liu et al: “ A Novel Antibody Engineering Strategy for Making Monovalent Bispecific Heterodimeric IgG Antibodies by Electro Related U . S . Application Data static Steering Mechanism ” , Journal of Biological Chemistry , vol . 290 , No. 12 , pp . 7535 - 7562 , 2015 . 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CI. 053537 , dated Dec . 2 , 2015 ; 23 pages. CPC . .. CO7K 16 / 2863 ( 2013 .01 ) ; A61P 27 / 16 ( Continued ) ( 2018 . 01 ) ; CO7K 16 /00 (2013 . 01 ) ; CO7K 16 / 24 ( 2013 . 01 ) ; C07K 16 / 244 (2013 . 01 ) ; COZK Primary Examiner - Sean E Aeder 2317 / 24 ( 2013 .01 ) ; CO7K 2317/ 31 ( 2013 . 01 ) ; CO7K 2317/ 35 (2013 .01 ) ; CO7K 2317/ 522 (57 ) ABSTRACT ( 2013 .01 ) ; COZK 2317 / 55 (2013 .01 ) ; COOK The present invention relates to engineered heteromultim eric proteins , and more specifically , to methods for produc 2317/ 75 ( 2013 .01 ); CO7K 2317/ 92 (2013 . 01 ) ing and purifying heterodimeric proteins, such as bispecific (58 ) Field of Classification Search antibodies and . Methods for producing and purifying such None engineered heterodimeric proteins and their use in diagnos See application file for complete search history. tics and therapeutics are also provided . 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