Affinity-Based Methods for Site-Specific Conjugation of Antibodies

Abstract

Conjugation of various reagents to antibodies has long been an elegant way to combine the superior binding features of the antibody with other desired but non-natural functions. Applications range from labels for detection in different analytical assays to the creation of new drugs by conjugation to molecules which improves the pharmaceutical effect. In many of these applications, it has been proven advantageous to control both the site and the stoichiometry of the conjugation to achieve a homogeneous product with predictable, and often also improved, characteristics. For this purpose, many research groups have, during the latest decade, reported novel methods and techniques, based on small molecules, peptides, and proteins with inherent affinity for the antibody, for site-specific conjugation of antibodies. This review provides a comprehensive overview of these methods and their applications and also describes a historical perspective of the field.

Template directed synthesis of antibody Fc conjugates with concomitant ligand release

“…Template directed ligation is emerging as powerful means to site specically label unmodied antibodies. 29,30 Through templating the chemical reaction with an aptamer, 31 a protein, 32 a small molecule 33 or a peptide, 34,35 authors were able to regioselectively label unmodied proteins. However, several drawbacks still remain to be solved because of the need to use UV irradiation, possible immunogenicity due to the covalent linkage of the protein or peptide ligand, or additional chemical steps to remove the ligand.…”

Enhanced susceptibility of SARS-CoV-2 spike RBD protein assay targeted by cellular receptors ACE2 and CD147: Multivariate data analysis of multisine impedimetric response

“…The control over the position and orientation of IgG2B during the covalent binding at the sensor’s surface is a challenge. Most widely used conjugation/labelling methods do not provide sufficient control over the label’s exact position, therefore, the optimal orientation of the IgG2B cannot be guaranteed [77] . This, in turn, can pose problems with a significant decrease in the antigen-binding activity.…”

Enhanced susceptibility of SARS-CoV-2 spike RBD protein assay targeted by cellular receptors ACE2 and CD147: multivariate data analysis of multisine impedimetric response

“…The control over the position and orientation of IgG2B during the covalent binding at the sensor's surface is a challenge. Most widely used conjugation/labelling methods do not provide sufficient control over the label's exact position, therefore, the optimal orientation of the IgG2B cannot be guaranteed (von Witting et al, 2021). This, in turn, can pose problems with a significant decrease in the antigen-binding activity.…”