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

Abstract

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 antibodiesBasic Protocol 2: PARP1 auto‐modification and antibody conjugation