Suppressing allostery in epitope mapping experiments using millisecond hydrogen / deuterium exchange mass spectrometry.

ABSTRACT: Localization of the interface between the candidate antibody and its antigen target, commonly known as epitope mapping, is a critical component of the development of therapeutic monoclonal antibodies. With the recent availability of commercial automated systems, hydrogen / deuterium eXchange (HDX) is rapidly becoming the tool for mapping epitopes preferred by researchers in both industry and academia. However, this approach has a significant drawback in that it can be confounded by 'allosteric' structural and dynamic changes that result from the interaction, but occur far from the point(s) of contact. Here, we introduce a 'kinetic' millisecond HDX workflow that suppresses allosteric effects in epitope mapping experiments. The approach employs a previously introduced microfluidic apparatus

SUBMITTER: Deng B 

PROVIDER: S-EPMC5680795 | biostudies-literature | 2017 Nov/Dec

REPOSITORIES: biostudies-literature