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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 established C-terminal cleavage method of cyanogen bromide in our laboratory and the optimized mass spectrometry, eight recombinant protein drugs with C terminal length of 2 ~ 37 amino acids and relative molecular mass of 200 ~ 5000 Test. (2) Mass spectrometry, positive ion mode, the atomization gas is nitrogen, the collision gas is Argon. Source temperature 80 ℃, cone voltage 50 V, MCP detector voltage of 2.15 kV. Results: The C-terminal of all eight recombinant protein drugs were successfully detected and basically the base peak. Conclusion: It is of high practical value and academic significance to establish a C-terminal sequencing recombinant protein drug for the detection of real drugs.