Cysteine Re-bridging Using Divinyl Pyrimidine Reagents for the Generation of Antibody-Drug Conjugates.

Antibody-drug conjugates (ADCs) are a powerful form of targeted therapy that can deliver drugs with a high level of selectivity towards a specific cell type, reducing off-target effects and increasing the therapeutic window compared to small-molecule therapeutics. However, creating ADCs that are stable, homogeneous, and with a controlled drug-to-antibody ratio (DAR) remains a significant challenge. Divinyl pyrimidine (DVP) reagents were developed as a method to irreversibly conjugate linker-payloads onto native antibodies to generate stable ADCs and antibody-probe systems with a precise drug-to-antibody ratio (DAR) of 4. This protocol describes how to conjugate DVPs to antibodies and methods for ADC characterization.