Guiding bispecific monovalent antibody formation through proteolysis of IgG1 single-chain. Issue 3 (3rd April 2017)

• Record Type: Journal Article
• Authors: Dimasi, Nazzareno; Fleming, Ryan; Sachsenmeier, Kris F.; Bezabeh, Binyam; Hay, Carl; Wu, Jincheng; Sult, Erin; Rajan, Saravanan; Zhuang, Li; Cariuk, Peter; Buchanan, Andrew; Bowen, Michael A.; Wu, Herren; Gao, Changshou
• Is Part Of: MAbs. Volume 9:Issue 3(2017)
• Page Start: 438
• Page End: 454
• Publication Date: 2017-04-03
• Subjects: Antibody engineering -- bispecific monovalent antibodies -- dual targeting -- single-chain antibody -- tethered single-chain antibodies; Monoclonal antibodies -- Therapeutic use -- Periodicals; Monoclonal antibodies -- Periodicals; Antibodies, Monoclonal -- Periodicals; 616.0798
• DOI: 10.1080/19420862.2016.1277301
• Languages: English
• ISSNs: 1942-0862

Abstract

ABSTRACT: We developed an IgG1 domain-tethering approach to guide the correct assembly of 2 light and 2 heavy chains, derived from 2 different antibodies, to form bispecific monovalent antibodies in IgG1 format. We show here that assembling 2 different light and heavy chains by sequentially connecting them with protease-cleavable polypeptide linkers results in the generation of monovalent bispecific antibodies that have IgG1 sequence, structure and functional properties. This approach was used to generate a bispecific monovalent antibody targeting the epidermal growth factor receptor and the type I insulin-like growth factor receptor that: 1) can be produced and purified using standard IgG1 techniques; 2) exhibits stability and structural features comparable to IgG1; 3) binds both targets simultaneously; and 4) has potent anti-tumor activity. Our strategy provides new engineering opportunities for bispecific antibody applications, and, most importantly, overcomes some of the limitations (e.g., half-antibody and homodimer formation, light chains mispairing, multi-step purification), inherent with some of the previously described IgG1-based bispecific monovalent antibodies.