High-Throughput Generation of Bipod (Fab x scFv) Bispecific Antibodies Exploits Differential Chain Expression and Affinity Capture

被引：18

作者：

Nesspor, Thomas C. ^{[1
]}

Kinealy, Kyle ^{[1
]}

Mazzanti, Nicholas ^{[1
]}

Diem, Michael D. ^{[1
]}

Boye, Kevin ^{[1
]}

Hoffman, Hunter ^{[1
]}

Springer, Christine ^{[1
]}

Sprenkle, Justin ^{[1
]}

Powers, Gordon ^{[1
]}

Jiang, Haiyan ^{[1
]}

La Porte, Sherry L. ^{[1
]}

Ganesan, Rajkumar ^{[1
]}

Singh, Sanjaya ^{[1
]}

Zwolak, Adam ^{[1
]}

机构：

[1] Janssen Res & Dev LLC, Biol Discovery, Spring House, PA 19477 USA

来源：

SCIENTIFIC REPORTS | 2020年 / 10卷 / 01期

关键词：

IGG ANTIBODIES; LIGHT-CHAIN; DESIGN;

D O I：

10.1038/s41598-020-64536-w

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Generation of bispecific antibodies (BsAbs) having two unique Fab domains requires heterodimerization of the two heavy chains and pairing of each heavy chain with its cognate light chain. An alternative bispecific scaffold (Bipod) comprising an scFv and a Fab on a heterodimeric Fc eliminates the possibility of light chain mispairing. However, unpredictable levels of chain expression and scFv-induced aggregation can complicate purification and reduce the yield of desired Bipod. Here, we describe a high-throughput method for generation of Bipods based on protein A and CH1 domain affinity capture. This method exploits over-expression of the scFv chain to maximize heterodimer yield. Bipods purified by this method have purity suitable for cell-based functional assays and in vivo studies.

引用

页数：10

共 21 条

[1] Bispecific Antibodies and Antibody-Drug Conjugates (ADCs) Bridging HER2 and Prolactin Receptor Improve Efficacy of HER2 ADCs [J].

Andreev, Julian ;

Thambi, Nithya ;

Bay, Andres E. Perez ;

Delfino, Frank ;

Martin, Joel ;

Kelly, Marcus P. ;

Kirshner, Jessica R. ;

Rafique, Ashique ;

Kunz, Arthur ;

Nittoli, Thomas ;

MacDonald, Douglas ;

Daly, Christopher ;

Olson, William ;

Thurston, Gavin .

MOLECULAR CANCER THERAPEUTICS, 2017, 16 (04) :681-693

[2] The making of bispecific antibodies [J].

Brinkmann, Ulrich ;

Kontermann, Roland E. .

MABS, 2017, 9 (02) :182-212

[3] Recombinant Expression of the Full-length Ectodomain of LDL Receptor-related Protein 1 (LRP1) Unravels pHdependent Conformational Changes and the Stoichiometry of Binding with Receptor- associated Protein ( RAP) [J].

De Nardis, Camilla ;

Lossl, Philip ;

van den Biggelaar, Maartje ;

Madoori, Pramod K. ;

Leloup, Nadia ;

Mertens, Koen ;

Heck, Albert J. R. ;

Gros, Piet .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2017, 292 (03) :912-924

[4] Efficient production of bispecific IgG of different isotypes and species of origin in single mammalian cells [J].

Dillon, Michael ;

Yin, Yiyuan ;

Zhou, Jianhui ;

McCarty, Luke ;

Ellerman, Diego ;

Slaga, Dionysos ;

Junttila, Teemu T. ;

Han, Guanghui ;

Sandoval, Wendy ;

Ovacik, Meric A. ;

Lin, Kedan ;

Hu, Zhilan ;

Shen, Amy ;

Corn, Jacob E. ;

Spiess, Christoph ;

Carter, Paul J. .

MABS, 2017, 9 (02) :213-230

[5] Considerations for the Design of Antibody-Based Therapeutics [J].

Goulet, Dennis R. ;

Atkins, William M. .

JOURNAL OF PHARMACEUTICAL SCIENCES, 2020, 109 (01) :74-103

[6] Fc-mediated activity of EGFR x c-Met bispecific antibody JNJ-61186372 enhanced killing of lung cancer cells [J].

Grugan, Katharine D. ;

Dorn, Keri ;

Jarantow, Stephen W. ;

Bushey, Barbara S. ;

Pardinas, Jose R. ;

Laquerre, Sylvie ;

Moores, Sheri L. ;

Chiu, Mark L. .

MABS, 2017, 9 (01) :114-126

[7] Immunoglobulin Fc Heterodimer Platform Technology: From Design to Applications in Therapeutic Antibodies and Proteins [J].

Ha, Ji-Hee ;

Kim, Jung-Eun ;

Kim, Yong-Sung .

FRONTIERS IN IMMUNOLOGY, 2016, 7

[8] Bispecific antibodies: a mechanistic review of the pipeline [J].

Labrijn, Aran F. ;

Janmaat, Maarten L. ;

Reichert, Janice M. ;

Parren, Paul W. H. I. .

NATURE REVIEWS DRUG DISCOVERY, 2019, 18 (08) :585-608

[9] Efficient generation of stable bispecific IgG1 by controlled Fab-arm exchange [J].

Labrijn, Aran F. ;

Meesters, Joyce I. ;

de Goeij, Bart E. C. G. ;

van den Bremer, Ewald T. J. ;

Neijssen, Joost ;

van Kampen, Muriel D. ;

Strumane, Kristin ;

Verploegen, Sandra ;

Kundu, Amitava ;

Gramer, Michael J. ;

van Berkel, Patrick H. C. ;

van de Winkel, Jan G. J. ;

Schuurman, Janine ;

Parren, Paul W. H. I. .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2013, 110 (13) :5145-5150

[10] Generation of bispecific IgG antibodies by structure-based design of an orthogonal Fab interface [J].

Lewis, Steven M. ;

Wu, Xiufeng ;

Pustilnik, Anna ;

Sereno, Arlene ;

Huang, Flora ;

Rick, Heather L. ;

Guntas, Gurkan ;

Leaver-Fay, Andrew ;

Smith, Eric M. ;

Ho, Carolyn ;

Hansen-Estruch, Christophe ;

Chamberlain, Aaron K. ;

Truhlar, Stephanie M. ;

Conner, Elaine M. ;

Atwell, Shane ;

Kuhlman, Brian ;

Demarest, Stephen J. .

NATURE BIOTECHNOLOGY, 2014, 32 (02) :191-+

← 1 2 3 →