基于质谱技术的贝伐珠单抗及其糖基化修饰的表征、定量与药代动力学分析

引用本文: 丛宇婷, 胡良海, 叶明亮, 顾景凯, 邹汉法. 基于质谱技术的贝伐珠单抗及其糖基化修饰的表征、定量与药代动力学分析[J]. 分析化学, 2017, 45(11): 1678-1685. doi: 10.11895/j.issn.0253-3820.170170 shu

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

基于质谱技术的贝伐珠单抗及其糖基化修饰的表征、定量与药代动力学分析

1.
吉林大学生命科学学院, 长春 130012;
2.
中国科学院大连化学物理研究所, 中国科学院分离分析化学重点实验室, 大连 116023
基金项目:
本文系国家自然科学基金项目（No.81373374）资助

摘要: 利用基质辅助激光解析电离飞行时间质谱技术、SDS-聚丙烯酰胺凝胶电泳技术以及Shot-gun蛋白质组学策略对贝伐珠单克隆抗体药物及其糖基化修饰进行了全面表征，共发现17种糖型并确定了特征肽段序列。利用液相色谱-串联质谱联用技术，采用平行反应检测模式，通过测定单抗水解后产生的特征肽段，实现了对大鼠血浆样品中单抗药物含量的绝对定量分析，获得不同给药计量下的贝伐珠单抗的药物浓度-时间曲线，同时实现了对单抗糖基化修饰各糖型的相对定量分析。本实验首先建立标准工作曲线，对大鼠血浆样品中的贝伐珠单抗进行定量分析，在线性范围内，相关系数R²=0.998，定量限为66 fmol，线性关系良好。对大鼠血浆样品的测定结果表明，高、低计量给药的贝伐珠单抗药物浓度-时间曲线趋势基本一致，浓度均为直接下降，与理论趋势相符。但是大多数糖型呈现浓度先上升的现象，之后的代谢情况因糖型差异而有所不同。

关键词:

English

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: 07 September 2017
Fund Project:
This work was supported by the National Natural Science Foundation of China (No. 81373374).

Abstract: 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.

Key words:

Bevacizumab
/ Glycosylation
/ Liquid chromatography-tandem mass spectrometry
/ Parallel reaction monitoring
/ Pharmacokinetics

1. [1]
  Chen Z, Wang J, Bao L, Guo L, Zhang W, Xue Y, Zhou H, Xiao Y, Wang J, Wu F, Deng Y, Qin C, Jin Q. Nat. Commun., 2015, 6:6714
2. [2]
  Hogarth P M, Pietersz G A. Nat. Rev. Drug Discov., 2012, 11(4):311-331
3. [3]
  Al-Debasi T, Al-Bekairy A, Al-Katheri A, Al Harbi S, Mansour M.Saudi J. Ophthalmol., 2017, 31(2):99-105
4. [4]
  Reusch D, Haberger M, Maier B, Maier M, Kloseck R, Zimmermann B, Hook M, Szabo Z, Tep S, Wegstein J, Alt N, Bulau P, Wuhrer M. MAbs, 2015, 7(1):167-179
5. [5]
  Beck A, Wagner-Rousset E, Ayoub D, Van Dorsselaer A, Sanglier-Cianferani S. Anal. Chem., 2013, 85(2):715-736
6. [6]
  Chung S, Quarmby V, Gao X,Ying Y, Lin L, Reed C, Fong C, Lau W, Qiu Z J, Shen A, Vanderlaan M, Song A. MAbs, 2012, 4(3):326-340
7. [7]
  Scallon B J,Tam S H, McCarthy S G, Cai A N, Raju T S. Mol. Immunol., 2007, 44(7):1524-1534
8. [8]
  Engvall E, Perlmann P. Immunochemistry, 1971, 8(9):871-874
9. [9]
  Gan S D, Patel K R. J. Invest. Dermatol., 2013, 133(9):e12
10. [10]
  Mesmin C, Fenaille F, Ezan E, Becher F. Bioanalysis, 2011, 3(5):477-480
11. [11]
  Ezan E, Bitsch F. Bioanalysis, 2009, 1(8):1375-1388
12. [12]
  Ronsein G E, Pamir N, von Haller P D, Kim D S, Oda M N, Jarvik G P, Vaisar T, Heinecke J W. J. Proteomics, 2015, 113:388-399
13. [13]
  Toyama A, Nakagawa H, Matsuda K, Sato TA, Nakamura Y, Ueda K. Anal. Chem., 2012, 84(22):9655-9662
14. [14]
  Hong Q, Lebrilla C B, Miyamoto S, Ruhaak L R. Anal. Chem., 2013, 85(18):8585-8593
15. [15]
  Lesur A, Domon B. Proteomics, 2015, 15(5-6):880-890
16. [16]
  Tsuchiya H, Tanaka K, Saeki Y. Biochem. Biophys. Res. Commun., 2013, 436(2):223-229
17. [17]
  Urisman A, Levin R S, Gordan J D, Webber J T, Hernandez H, Ishihama Y, Shokat K M, Burlingame A L. Mol. Cell Proteomics, 2017, 16(2):265-277
18. [18]
  Sundberg M, Strage E M, Bergquist J, Holst B S, Ramstr m M. PLos One, 2016, 11(12):e0167138
19. [19]
  Siemiatkoski J, Lyubarskaya Y, Houde D, Tep S, Mhatre R. Carbohydr. Res., 2006, 341(3):410-419
20. [20]
  Bobaly B, D'Atri V, Goyon A, Colas O, Beck A, Fekete S, Guillarme D. J. Chromatogr. B, 2017, 1060:325-335
21. [21]
  Cheng K, Chen R, Seebun D, Ye M, Figeys D, Zou H. J. Proteomics, 2014, 110:145-154

扫一扫看文章

计量

PDF下载量: 7
文章访问数: 753
HTML全文浏览量: 103

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

基于质谱技术的贝伐珠单抗及其糖基化修饰的表征、定量与药代动力学分析

English

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

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于质谱技术的贝伐珠单抗及其糖基化修饰的表征、定量与药代动力学分析

English

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

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs