Site-Specific Antibody Conjugation with Payloads beyond Cytotoxins

As antibody-drug conjugates have become a very important modality for cancer therapy, many site-specific conjugation approaches have been developed for generating homogenous molecules. The selective antibody coupling is achieved through antibody engineering by introducing specific amino acid or unnatural amino acid residues, peptides, and glycans. In addition to the use of synthetic cytotoxins, these novel methods have been applied for the conjugation of other payloads, including non-cytotoxic compounds, proteins/peptides, glycans, lipids, and nucleic acids. The non-cytotoxic compounds include polyethylene glycol, antibiotics, protein degraders (PROTAC and LYTAC), immunomodulating agents, enzyme inhibitors and protein ligands. Different small proteins or peptides have been selectively conjugated through unnatural amino acid using click chemistry, engineered C-terminal formylglycine for oxime or click chemistry, or specific ligation or transpeptidation with or without enzymes. Although the antibody protamine peptide fusions have been extensively used for siRNA coupling during early studies, direct conjugations through engineered cysteine or lysine residues have been demonstrated later. These site-specific antibody conjugates containing these payloads other than cytotoxic compounds can be used in proof-of-concept studies and in developing new therapeutics for unmet medical needs.

Automated summary

As antibody-drug conjugates have become crucial for cancer therapy, various approaches have been developed to achieve selective and homogeneous molecules. These methods involve engineering antibodies to introduce specific amino acids or peptides for coupling. In addition to cytotoxic compounds, these techniques can also be applied to conjugate non-cytotoxic compounds, proteins/peptides, glycans, lipids, and nucleic acids. The resulting site-specific antibody conjugates offer potential for proof-of-concept studies and the development of novel therapeutic options.