Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, California.
Department of Chemistry, University of Southern California, Los Angeles, California.
Curr Protoc. 2023 Dec;3(12):e958. doi: 10.1002/cpz1.958.
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. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Expression and purification of PARP1 and antibodies Basic Protocol 2: PARP1 auto-modification and antibody conjugation.
双特异性抗体在治疗开发方面引起了相当多的研究兴趣。已经建立了许多遗传和化学方法来生产具有不同形式的双特异性抗体。本方案描述了一种利用源自翻译后多聚 ADP-核糖基化的化学功能化聚 ADP-核糖聚合物合成双特异性抗体的新方法。基本方案 1 包括表达和纯化重组全长人多聚 ADP-核糖聚合酶 1 (PARP1) 以及针对 T 细胞 CD3 和乳腺癌肿瘤相关人表皮生长因子受体 2 (HER2) 分子的单克隆抗体的实验程序。基本方案 2 详细介绍了通过 PARP1 酶促制备功能化多聚 ADP-核糖聚合物和抗体分子化学偶联用于双特异性抗体生产的方法。© 2023 作者。Wiley Periodicals LLC 出版的《当代协议》。基本方案 1：PARP1 和抗体的表达和纯化 基本方案 2：PARP1 自动修饰和抗体偶联。
