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免脱盐柱除盐的蛋白组学前处理方法

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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.
Proteomic pretreatment method without desalination column
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 re-placed with ammonium bicarbonate solution by centrifugation before enzymatic digestion using ultrafil-tration 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.

proteomic analysissample pre-treatmentdesalting columnheatingammonium bicar-bonate

季学猛、史爱莹、吴景、张燕、王硕

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南开大学 医学院,天津 300071

蛋白组学 样品前处理 脱盐柱 加热 碳酸氢铵

国家重点研发计划项目

32030083

2024

实验室科学
南开大学 中国高等教育学会

实验室科学

影响因子:0.805
ISSN:1672-4305
年,卷(期):2024.27(5)