Modulation of protein A binding allows single-step purification of mouse bispecific antibodies that retain FcRn binding
| Autor: | Anthony A. Armstrong, Eva Emmell, Dennis R. Goulet, Songmao Zheng, Kerry Brosnan, Jose Pardinas, Jeffrey Luo, Susan H. Tam, Mark L. Chiu, Gary L. Gilliland, Adam Zwolak |
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| Rok vydání: | 2017 |
| Předmět: |
Models
Molecular 0301 basic medicine medicine.drug_class Protein subunit Immunology Mutant Receptors Fc bispecific Crystallography X-Ray medicine.disease_cause Monoclonal antibody Mice 03 medical and health sciences 0302 clinical medicine Protein Domains Affinity chromatography Report antibody Antibodies Bispecific medicine Animals Humans Immunology and Allergy Staphylococcal Protein A x-ray crystallography Mutation biology Chemistry Histocompatibility Antigens Class I Antibodies Monoclonal Fragment crystallizable region 030104 developmental biology Biochemistry Immunoglobulin G 030220 oncology & carcinogenesis biology.protein Mutant Proteins Antibody Protein A calorimetry pharmacokinetics Protein Binding |
| Zdroj: | mAbs |
| ISSN: | 1942-0870 1942-0862 |
| DOI: | 10.1080/19420862.2017.1375639 |
| Popis: | The increased number of bispecific antibodies (BsAb) under therapeutic development has resulted in a need for mouse surrogate BsAbs. Here, we describe a one-step method for generating highly pure mouse BsAbs suitable for in vitro and in vivo studies. We identify two mutations in the mouse IgG2a and IgG2b Fc region: one that eliminates protein A binding and one that enhances protein A binding by 8-fold. We show that BsAbs harboring these mutations can be purified from the residual parental monoclonal antibodies in one step using protein A affinity chromatography. The structural basis for the effects of these mutations was analyzed by X-ray crystallography. While the mutation that disrupted protein A binding also inhibited FcRn interaction, a bispecific mutant in which one subunit retained the ability to bind protein A could still interact with FcRn. Pharmacokinetic analysis of the serum half-lives of the mutants showed that the mutant BsAb had a serum half-life comparable to a wild-type Ab. The results describe a rapid method for generating panels of mouse BsAbs that could be used in mouse studies. |
| Databáze: | OpenAIRE |
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