重组蛋白药物C末端不同长度氨基酸序列的质谱分析

目的: 基于本实验室已建立的溴化氰裂解蛋白质C末端方法结合优化后的质谱检测技术,对C末端长度分别为2~37个氨基酸,相对分子质量在200~5000的8个重组蛋白药物进行检测。 方法: (1)针对重组蛋白药物的不同状态(SDS-PAGE、干粉或溶液)分别进行C末端胶内或溶液裂解;(2)质谱检测,正离子方式,雾化气为氮气,碰撞气体为氩气。源温80 ℃,锥孔电压50 V,MCP检测器电压为2.15 kV。 结果: 8个重组蛋白药物的C末端全部成功检测出,且基本为基峰。 结论: 建立的重组蛋白药物C末端测序联用方法应用于实际药物的检测具有很高的实用价值和学术意义。

Objective: Based on the method established by cyanogen bromide cleavage of proteins C-terminal combined with the optimized mass spectrometry in our laboratory,detection of C-terminal lengths of 2 to 37 amino acids,relative molecular mass 200-5000 of the 8 recombinant protein drugs. Methods: (1) For different states of recombinant protein drugs (SDS-PAGE,dry powder or solution) to C terminal cleavage in gel or solution,respectively.(2) Mass spectrometry detection,positive ion mode,atomization gas was nitrogen,collision gas was argon,source temperature 80 ℃,cone voltage 50 V,MCP detector voltage of 2.15 kV. Results: All of C-terminal of the 8 recombinant protein drugs successfully detected as the base peak. Conclusions: The established C-terminal sequencing method of the recombinant protein was applied to the actual drugs testing,have high practical value and academic significance.