Analysis of therapeutic monoclonal antibody glycoforms by mass spectrometry for pharmacokinetics study.
Monoclonal antibodies (mAbs), are one of the most important protein drugs have attracted increasing attention. However, the pharmacokinetics of mAbs has not been fully investigated due to the complexity of protein drugs. Traditonal immuno-based approaches can not recognize the proteoforms of mAbs because of the long development cycles, prohibitive cost, and interactions between different proteins. Therefore, reliable qualitative and quantitative analysis of the proteoforms of mAbs in biological samples is of crucial importance. Herein, a novel method was developed for absolute quantitation of mAbs and their glycoforms in complex biological samples such as serum and tissues. With the combination of HILIC enrichment and parallel reaction monitoring by high resolution mass spectrometry, most of the glycoforms can be accurately quantified at the fmol level through the use of the model mAb of bevacizumab. More importantly, the structural confirmation can be achieved simultaneously without the need for additional experiments. This strategy can be readily applied to the pharmacokinetic study of glycosylation modification and biomarker discovery for clinical applications.
School of Life Sciences, Jilin University, Changchun 130012, China; CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
Yuting
Y
Cong
School of Life Sciences, Jilin University, Changchun 130012, China. Electronic address: lianghaihu@jlu.edu.cn.
Lianghai
L
Hu
CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
Zhang
Z
Zhang
School of Life Sciences, Jilin University, Changchun 130012, China.
Yin
Y
Gao
CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
Mingming
M
Dong
CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
Hongqiang
H
Qin
CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. Electronic address: mingliang@dicp.ac.cn.
Mingliang
M
Ye
School of Life Sciences, Jilin University, Changchun 130012, China.
Jingkai
J
Gu
CAS Key Laboratory of Separation Sciences for Analytical Chemistry, National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
Hanfa
H
Zou
D000074322:Antineoplastic Agents, Immunological / Q000032:analysis* / Q000737:chemistry*; D000068258:Bevacizumab / Q000032:analysis* / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D013058:Mass Spectrometry / Q000379:methods*; D011499:Protein Processing, Post-Translational
D000074322:Antineoplastic Agents, Immunological; D007074:Immunoglobulin G; D000068258:Bevacizumab
Glycosylation; Mass spectrometry; Parallel reaction monitoring; Pharmacokinetics; Proteomics; Therapeutic monoclonal antibody