Liquid-phase separations coupled with ion mobility-mass spectrometry for next-generation biopharmaceutical analysis

ABSTRACT

Introduction

The pharmaceutical industry continues to expand its search for innovative biotherapeutics. The comprehensive characterization of such therapeutics requires many analytical techniques to fully evaluate critical quality attributes, making analysis a bottleneck in discovery and development timelines. While thorough characterization is crucial for ensuring the safety and efficacy of biotherapeutics, there is a need to further streamline analytical characterization and expedite the overall timeline from discovery to market.

Areas covered

This review focuses on recent developments in liquid-phase separations coupled with ion mobility-mass spectrometry (IM-MS) for the development and characterization of biotherapeutics. We cover uses of IM-MS to improve the characterization of monoclonal antibodies, antibody-drug conjugates, host cell proteins, glycans, and nucleic acids. This discussion is based on an extensive literature search using Web of Science, Google Scholar, and SciFinder.

Expert opinion

IM-MS has the potential to enhance the depth and efficiency of biotherapeutic characterization by providing additional insights into conformational changes, post-translational modifications, and impurity profiles. The rapid timescale of IM-MS positions it well to enhance the information content of existing assays through its facile integration with standard liquid-phase separation techniques that are commonly used for biopharmaceutical analysis.

KEYWORDS:

• Biotherapeutics
• ion mobility
• mass spectrometry
• proteins
• separations

Article highlights

• Ion mobility improves proteomic peak capacity and depth of analysis.

• High-resolution ion mobility technologies such as structures for lossless ion manipulations and cyclic ion mobility rapidly separates isomeric peptides.

• Ion mobility can be integrated with nondenaturing separation methods to provide more detailed insight into therapeutic protein structure, dynamics, and interactions.

• Collision induced unfolding can be used with ion mobility to provide information about both structure and stability of proteins following nondenaturing liquid-phase separations.

Declaration of interest

The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

Reviewer disclosures

Peer reviewers on this manuscript have no relevant financial or other relationships to disclose.

Additional information

Funding

This work was supported by funding from the National Science Foundation Graduate Research Fellowship Program under grant number DGE-1841052 (D.M.M.). The development of IM-MS technologies for applications in the pharmaceutical industry is supported in the Ruotolo lab by the National Institutes of Health (R01 GM095832, RM1 GM149374), the National Science Foundation (2304961), and Agilent Technologies (to B.T.R.).