Affiliation:
1. Department of Pharmacology and Pharmaceutical Sciences University of Southern California Los Angeles California
2. Department of Chemistry University of Southern California Los Angeles California
3. Norris Comprehensive Cancer Center University of Southern California Los Angeles California
4. Research Center for Liver Diseases University of Southern California Los Angeles California
Abstract
AbstractBispecific 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
Funder
National Institute of General Medical Sciences
National Institutes of Health
National Institute of Biomedical Imaging and Bioengineering
National Cancer Institute
National Institute of Diabetes and Digestive and Kidney Diseases
Subject
Medical Laboratory Technology,Health Informatics,General Pharmacology, Toxicology and Pharmaceutics,General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Neuroscience