Plasma Induced Modification of Biomolecules (PLIMB) for Epitope Mapping

Abstract

Epitope mapping is the process of identifying the site(s) of an antigen where an antibody binds. Knowledge of epitopes facilitates drug design and patent applications; hence it is a key element of vaccine and drug development. Hydroxyl radical protein footprinting coupled with mass spectrometry offers opportunities to map antibody-binding regions of antigens. We report here the use of hydroxyl radical protein footprinting via Plasma Induced Modification of Biomolecules (PLIMB) to characterize the epitopes of antithrombin bound to the serine-protease thrombin, and monoclonal antibody (mAb) IIB8 to human transcription factor II B (TFIIB). The protein samples were exposed to plasma for hydroxyl radical labeling and the labeled sites were identified using mass spectrometry. The data was analyzed using Byos (Protein Metrics). The PLIMB data is compared to previously published results that determined the epitope of thrombin and TFIIB using other techniques.