Qualitative and quantitative characterization of protein biotherapeutics with liquid chromatography mass spectrometry.
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. © 2016 Wiley Periodicals, Inc. Mass Spec Rev 36:734-754, 2017.
Beijing University of Chinese Medicine, Beijing, 100029, China.|Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, NY, 14214.|New York State Center of Excellence in Bioinformatics and Life Sciences, Buffalo, NY, 14203.
Miao
M
Qu
Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, NY, 14214.|New York State Center of Excellence in Bioinformatics and Life Sciences, Buffalo, NY, 14203.
Bo
B
An
Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, NY, 14214.|New York State Center of Excellence in Bioinformatics and Life Sciences, Buffalo, NY, 14203.
Shichen
S
Shen
Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, NY, 14214.|New York State Center of Excellence in Bioinformatics and Life Sciences, Buffalo, NY, 14203.
Ming
M
Zhang
Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, NY, 14214.|New York State Center of Excellence in Bioinformatics and Life Sciences, Buffalo, NY, 14203.
Xiaomeng
X
Shen
State Key Laboratory of Toxicology and Medical Countermeasures, Beijing Institute of Pharmacology and Toxicology, Beijing, 100850, China.
Xiaotao
X
Duan
Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, NY, 14214.
Joseph P
JP
Balthasar
Department of Pharmaceutical Sciences, School of Pharmacy and Pharmaceutical Sciences, University at Buffalo, State University of New York, Buffalo, NY, 14214.|New York State Center of Excellence in Bioinformatics and Life Sciences, Buffalo, NY, 14203.
Jun
J
Qu
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000627:therapeutic use; D001688:Biological Products / Q000032:analysis; D002853:Chromatography, Liquid / Q000379:methods*; D004220:Disulfides / Q000032:analysis / Q000737:chemistry; D055808:Drug Discovery / Q000379:methods; D006031:Glycosylation; D006801:Humans; D018796:Immunoconjugates / Q000032:analysis; D013058:Mass Spectrometry / Q000379:methods*; D010449:Peptide Mapping / Q000379:methods; D011506:Proteins / Q000032:analysis* / Q000378:metabolism / Q000493:pharmacokinetics; D011994:Recombinant Proteins / Q000032:analysis; D012680:Sensitivity and Specificity; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D001688:Biological Products; D004220:Disulfides; D018796:Immunoconjugates; D011506:Proteins; D011994:Recombinant Proteins
antibody-drug conjugate; biotherapeutics; liquid chromatography; mass spectrometry; quantification