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Mass Spectrom Rev. 2017 Nov;36(6):734-754. doi: 10.1002/mas.21500. Epub 2016 Apr 20.

Qualitative and quantitative characterization of protein biotherapeutics with liquid chromatography mass spectrometry.

Qu M1,2,3, An B2,3, Shen S2,3, Zhang M2,3, Shen X2,3, Duan X4, Balthasar JP2, Qu J2,3.

Author information

1
Beijing University of Chinese Medicine, Beijing, 100029, China.
2
Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, NY, 14214.
3
New York State Center of Excellence in Bioinformatics and Life Sciences, Buffalo, NY, 14203.
4
State Key Laboratory of Toxicology and Medical Countermeasures, Beijing Institute of Pharmacology and Toxicology, Beijing, 100850, China.

Abstract

In the last decade, the advancement of liquid chromatography mass spectrometry (LC/MS) techniques has enabled their broad application in protein characterization, both quantitatively and qualitatively. Owing to certain important merits of LC/MS techniques (e.g., high selectivity, flexibility, and rapid method development), LC/MS assays are often deemed as preferable alternatives to conventional methods (e.g., ligand-binding assays) for the analysis of protein biotherapeutics. At the discovery and development stages, LC/MS is generally employed for two purposes absolute quantification of protein biotherapeutics in biological samples and qualitative characterization of proteins. For absolute quantification of a target protein in bio-matrices, recent work has led to improvements in the efficiency of LC/MS method development, sample treatment, enrichment and digestion, and high-performance low-flow-LC separation. These advances have enhanced analytical sensitivity, specificity, and robustness. As to qualitative analysis, a range of techniques have been developed to characterize intramolecular disulfide bonds, glycosylation, charge variants, primary sequence heterogeneity, and the drug-to-antibody ratio of antibody drug conjugate (ADC), which has enabled a refined ability to assess product quality. In this review, we will focus on the discussion of technical challenges and strategies of LC/MS-based quantification and characterization of biotherapeutics, with the emphasis on the analysis of antibody-based biotherapeutics such as monoclonal antibodies (mAbs) and ADCs.

KEYWORDS:

antibody-drug conjugate; biotherapeutics; liquid chromatography; mass spectrometry; quantification

PMID:
27097288
DOI:
10.1002/mas.21500
[Indexed for MEDLINE]

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