Anal Bioanal Chem DOI 10.1007/s00216-014-7924-3

REVIEW

Higher order structure characterization of protein therapeutics by hydrogen/deuterium exchange mass spectrometry Richard Y.-C. Huang & Guodong Chen

Received: 14 April 2014 / Revised: 18 May 2014 / Accepted: 22 May 2014 # Springer-Verlag Berlin Heidelberg 2014

Abstract Characterization of therapeutic drugs is a crucial step in drug development in the biopharmaceutical industry. Analysis of protein therapeutics is a challenging task because of the complexities associated with large molecular size and 3D structures. Recent advances in hydrogen/deuteriumexchange mass spectrometry (HDX-MS) have provided a means to assess higher-order structure of protein therapeutics in solution. In this review, the principles and procedures of HDX-MS for protein therapeutics characterization are presented, focusing on specific applications of epitope mapping for protein–protein interactions and higher-order structure comparison studies for conformational dynamics of protein therapeutics. Keywords Protein therapeutics . Higher-order structure . Mass spectrometry . HDX-MS

Introduction Therapeutic drugs span a wide range of molecular sizes from small molecules (