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

发布时间：2018-02-28 12:08

本文关键词： 贝伐珠单抗糖基化修饰液相色谱-串联质谱联用平行反应监测药代动力学　出处：《分析化学》2017年11期 　论文类型：期刊论文

【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。
[Abstract]:Using matrix assisted laser desorption ionization time of flight mass spectrometry and SDS- polyacrylamide gel electrophoresis and Shot-gun proteomics strategy, the bevacious monoclonal antibody drugs and their glycosylation modification were characterized. A total of 17 carbohydrates were found and the characteristic peptide segments were sequenced. By using liquid chromatography-tandem mass spectrometry (LC-MS) and parallel reaction detection mode, the characteristic peptides produced by hydrolysis of McAbs were determined. The absolute quantitative analysis of the content of monoclonal antibodies in rat plasma samples was achieved, and the concentration-time curves of bevacizumab in different dosages were obtained. At the same time, the relative quantitative analysis of each sugar type modified by glycosylation of monoclonal antibody was realized. In this experiment, the standard working curve was established, and the bevacizumab in rat plasma was quantitatively analyzed in a linear range. The correlation coefficient was 0.998, the quantitative limit was 66fmol, and the linear relationship was good. The results of determination of plasma samples from rats showed that the concentration-time curves of bevacizumab with high and low doses were basically the same, and the concentrations were all decreased directly. But the concentration of most sugars increased first, and then the metabolism was different according to the difference of sugar type.
【作者单位】：吉林大学生命科学学院;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;
【基金】：国家自然科学基金项目(No.81373374)资助~~
【分类号】：O657.63;TQ450.1

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