免脱盐柱除盐的蛋白组学前处理方法  

Proteomic pretreatment method without desalination column

作　　者：季学猛 史爱莹 吴景 张燕 王硕[1] JI Xuemeng;SHI Aiying;WU Jing;ZHANG Yan;WANG Shuo(School of Medicine,Nankai University,Tianjin 300071,China)

机构地区：[1]南开大学医学院,天津300071

出　　处：《实验室科学》2024年第5期33-35,40,共4页Laboratory Science

基　　金：国家重点研发计划项目(项目编号:32030083)。

摘　　要：现有的蛋白组样品前处理中脱盐柱除盐的方法存在步骤繁多、易损失微量样品等缺点,这里,一种超滤管辅助酶解及加热潮解除盐的蛋白组学前处理方法被提出。具体而言,是利用超滤管辅助,通过离心在酶解前将缓冲液置换成碳酸氢铵溶液,酶解后多肽溶液中的碳酸氢铵被加热除去,最终得到脱盐的纯净多肽。该方法操作简便,不损失样品,而且无需洗脱步骤,可以方便地与LC-MS直接衔接,进行质谱分析。The existing method of desalting column in proteomic sample pre-treatment has many steps and is prone to loss of trace samples.Here,a proteomic pre-treatment method of enzymatic digestion assisted by ultrafiltration tube and desalting by heating is proposed.Specifically,the buffer was replaced with ammonium bicarbonate solution by centrifugation before enzymatic digestion using ultrafiltration tube assistance,and the ammonium bicarbonate in the peptide solution was removed by heating after enzymatic digestion to finally obtain the desalted pure peptides.This method is easy to operate without loss of sample and does not require an elution step,and can be easily interfaced directly with LC-MS for further mass spectrometry analysis.

关 键 词：蛋白组学 样品前处理 脱盐柱 加热 碳酸氢铵 

分 类 号：G482[文化科学—教育学]