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Citation: CONG Yu-Ting, HU Liang-Hai, YE Ming-Liang, GU Jing-Kai, ZOU Han-Fa. Characterization, Quantification and Pharmacokinetic Analysis of Bevacizumab and Its Glycosylation by Mass Spectrometry[J]. Chinese Journal of Analytical Chemistry, ;2017, 45(11): 1678-1685. doi: 10.11895/j.issn.0253-3820.170170 shu

Characterization, Quantification and Pharmacokinetic Analysis of Bevacizumab and Its Glycosylation by Mass Spectrometry

1.
College of Life Science, Jilin University, Changchun 130012, China;
2.
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

Received Date: 20 March 2017
Revised Date: 7 September 2017

Fund Project: This work was supported by the National Natural Science Foundation of China (No. 81373374).

The bevacizumab and its glycoforms were analyzed by matrix-assisted laser desorption ionization-time of flight-time of flight-mass spectrometry (MALDI-TOF-MS), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and short-gun strategy, with the sequence of unique peptide and seventeen glycoforms being characterized. The bevacizumab and its glycopeptides concentrations in mice plasma with different intravenous injection doses of bevacizumab were detected and the concentration-time curves were obtained by parallel reaction monitoring (PRM) method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) technique. First, standard curves were created for quantification of mAb in mice plasma, which showed good linearity, with the correlation coefficient (R²) value of 0.998 and the lower limit of quantification of 66 fmol. Detection results of high and low doses of the drug in the mice plasma samples showed that the drug concentration-time curve trend was consistent, e.g. the concentration was decreasing. However, the results of quantitation of seventeen glycoforms demonstrated that the metabolism of different glycoforms was different. The concentrations of most glycoforms increased first, whereas the metabolism afterwards differed by different glycoforms.
- Bevacizumab,
- Glycosylation,
- Liquid chromatography-tandem mass spectrometry,
- Parallel reaction monitoring,
- Pharmacokinetics
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