title: Kinetic analysis of a monoclonal therapeutic antibody and its single-chain homolog by surface plasmon resonance.
abstract: Monoclonal antibodies (mAbs) and antibody fragments have become an emerging class of therapeutics since 1986. Their versatility enables them to be engineered for optimal efficiency and decreased immunogenicity, and the path to market has been set by recent regulatory approvals. One of the initial criteria for success of any protein or antibody therapeutic is to understand its binding characteristics to the target antigen. Surface plasmon resonance (SPR) has been widely used and is an important tool for ligand-antigen binding characterization. In this work, the binding kinetics of a recombinant mAb and its single-chain antibody homolog, single-chain variable fragment (scFv), was analyzed by SPR. These two proteins target the same antigen. The binding kinetics of the mAb (bivalent antibody) and scFv (monovalent scFv) for this antigen was analyzed along with an assessment of the thermodynamics of the binding interactions. Alternative binding configurations were investigated to evaluate potential experimental bias because theoretically experimental binding configuration should have no impact on binding kinetics. Self-association binding kinetics in the proteins' respective formulation solutions and antigen epitope mapping were also evaluated. Functional characterization of monoclonal and single-chain antibodies has become just as important as structural characterization in the biotechnology field.
authors.affiliation: Alexion Pharmaceuticals, Cheshire, CT 06410, USA. patelr@alxn.com
authors.forename: Rekha
authors.initials: R
authors.lastname: Patel
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Andrien
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism* / Q000627:therapeutic use*; D000941:Antigens / Q000378:metabolism; D018604:Epitope Mapping; D007135:Immunoglobulin Variable Region / Q000378:metabolism*; D007700:Kinetics; D017386:Sequence Homology, Amino Acid; D020349:Surface Plasmon Resonance; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D007135:Immunoglobulin Variable Region