Chapter 7 - Advanced structural elucidation of new antibody formats by mass spectrometry

Abstract

Boosted by the outstanding success of protein-based bio-therapeutics over the last decades, new product formats designed by sophisticated protein engineering enter progressively into the biopharmaceutical industry’s pipeline with major implications on the analytical methods applied for their characterization. While most of the critical quality attributes for standard (immunoglobulin G like) molecules can be adequately assessed and monitored by conventional approaches, the increasing complexity of novel formats requires adjustments to the analytical methodologies, especially for separation-based techniques coupled to ultraviolet/fluorescence detection. After introducing the current common practices for the characterization of biopharmaceuticals, this chapter then provides a review of emerging liquid chromatography–mass spectrometry (LC–MS)-based technologies, which have the potential to complement the preexisting analytical toolbox and improve product/process understanding. In this context, the benefits of native LC–MS and two-/multidimensional LC–MS applications to assess product attributes while preserving the higher order structure and chromatographic separation and detection resolution are discussed.