Generation of Bispecific Antibodies by Functionalized Poly-ADP-Ribose Polymers

被引：0
作者：
Kim, Hyo Sun [1 ]
Zhang, Yong [1 ,2 ,3 ,4 ]
机构：
[1] Univ Southern Calif, Dept Pharmacol & Pharmaceut Sci, Los Angeles, CA 90007 USA
[2] Univ Southern Calif, Dept Chem, Los Angeles, CA 90007 USA
[3] Univ Southern Calif, Norris Comprehens Canc Ctr, Los Angeles, CA 90007 USA
[4] Univ Southern Calif, Res Ctr Liver Dis, Los Angeles, CA 90007 USA
来源：
CURRENT PROTOCOLS | 2023年 / 3卷 / 12期
基金：
美国国家卫生研究院;
关键词：
antibody; bispecific antibody; chemical conjugation; immunotherapy; PARP1; poly-ADP-ribose; post-translational modification; POLY(ADP-RIBOSE) POLYMERASE; PARP; REPAIR; AUTOMODIFICATION; CANCER;
D O I：
10.1002/cpz1.958
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Bispecific antibodies have drawn considerate research interests for therapeutic development. Numerous genetic and chemical methods are established to produce bispecific antibodies with varied formats. This protocol describes a novel approach to the synthesis of bispecific antibodies by utilizing chemically functionalized poly-ADP-ribose polymers derived from post-translational poly-ADP-ribosylation. Basic Protocol 1 includes experimental procedures for expressing and purifying recombinant full-length human poly-ADP-ribose polymerase 1 (PARP1) as well as monoclonal antibodies targeting T-cell CD3 and breast cancer tumor-associated human epidermal growth factor receptor 2 (HER2) molecules. Basic Protocol 2 details methods for enzymatic preparation of functionalized poly-ADP-ribose polymers by PARP1 and chemical conjugation of antibody molecules for bispecific antibody production. (c) 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.Basic Protocol 1: Expression and purification of PARP1 and antibodiesBasic Protocol 2: PARP1 auto-modification and antibody conjugation
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页数：12
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