Journal Highlight: Qualitative and quantitative characterization of protein biotherapeutics with liquid chromatography mass spectrometry

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  • Published: Nov 6, 2017
  • Author: spectroscopyNOW
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thumbnail image: Journal Highlight: Qualitative and quantitative characterization of protein biotherapeutics with liquid chromatography mass spectrometry

<p>The technical challenges and strategies of LC/MS-based quantification and characterization of biotherapeutics have been reviewed, with emphasis on the analysis of antibody-based biotherapeutics such as monoclonal antibodies and antibody-drug conjugates.</p>

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

Mass Spectrometry Reviews, 2017, 36, 734-754
Miao Qu, Bo An, Shichen Shen, Ming Zhang, Xiaomeng Shen, Xiaotao Duan, Joseph P. Balthasar and Jun Qu

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.

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