Antibody conjugation via one and two C-terminal selenocysteines.

TitleAntibody conjugation via one and two C-terminal selenocysteines.
Publication TypeJournal Article
Year of Publication2013
AuthorsLi X, Yang J, Rader C
JournalMethods (San Diego, Calif.)
Date Published2013 Jun 10
Abstract

Conventional antibody conjugation methods generate antibody-drug conjugates that are heterogeneous mixtures with undefined stoichiometry and variable pharmacokinetic and pharmacodynamic properties. We have previously described a strategy to generate site-specific antibody conjugates by genetic engineering of an antibody with a single C-terminal selenocysteine, the 21st natural amino acid, which displays unique chemical reactivity allowing selective conjugation in the presence of all other natural amino acids. In the present work, we describe a method for expanding this technology to higher drug-to-antibody ratios by genetically engineering an antibody with two C-terminal selenocysteines. Both selenocysteines effectively conjugate to a fluorescent iodoacetamide derivative and the resulting conjugate fully retains its antigen binding capability. Our method provides a platform for creating stoichiometrically defined antibody-drug conjugates for therapeutic intervention.

DOI10.1155/2013/724183
Alternate JournalMethods