离心条件对丙酮沉淀法提取尿蛋白质的影响

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (2): 229-234.

离心条件对丙酮沉淀法提取尿蛋白质的影响

李京, 周冬冬, 孙伟^*

中国医学科学院基础医学研究所北京协和医学院基础学院药理系, 北京 100005

收稿日期:2019-10-12 修回日期:2019-12-20 出版日期:2020-02-05 发布日期:2020-02-05
通讯作者: ^*sunwei@ibms.pumc.edu.cn
基金资助:
国家科技部重点基础研究项目(2013FY114100);中国医学科学院医学科学创新基金(2017-I2M-1-009)

Effect of centrifugal conditions on urine protein extraction by acetone precipitation method

LI Jing, ZHOU Dong-dong, SUN Wei^*

Department of Pharmacology, Institute of Basic Medical Sciences of CAMS,School of Basic Medicine of PUMC, Beijing 100005, China

Received:2019-10-12 Revised:2019-12-20 Online:2020-02-05 Published:2020-02-05
Contact: ^*sunwei@ibms.pumc.edu.cn

摘要/Abstract

摘要： 目的探究丙酮沉淀后离心转速和离心温度对尿蛋白质组的影响及健康人尿液蛋白质组的个体差异。方法用不同离心转速(4 000×g和12 000×g)和离心温度(4 ℃和20 ℃)对丙酮沉淀后的人尿液样品进行处理,提取尿蛋白进行质谱分析,比较不同方法的鉴定效果,确定丙酮沉淀后尿蛋白离心的最佳条件。将上述方法用于36例(男女比例为1∶1)健康人尿液蛋白质组分析,研究健康人的个体差异。结果在低温高转速条件下(4 ℃,12 000×g),鉴定效果最好,健康人尿蛋白质组个体差异为0.635。结论丙酮沉淀后,低温高转速(4 ℃,12 000×g)提取尿蛋白的效果较好,尿蛋白的个体差异与血浆蛋白质组相近,具有一定的稳定性,可以用于大规模临床分析。

关键词: 离心温度和转速, 尿液蛋白质组学, 个体差异

Abstract: Objective To investigate the effect of centrifugal speed and temperature on urine proteome after acetone precipitation and the individual variation of urine proteome in normal people. Methods Human urine samples precipitated by acetone were centrifuged at different centrifugation speeds (4 000×g and 12 000×g) and temperatures (4 ℃ and 20 ℃). Urine proteins were extracted for mass spectrum analysis. The identification results of different methods were compared to determine the best conditions of urine protein centrifugation after acetone precipitation. The above methods were used to analyze the urine proteome of 36 normal people (male∶female =1∶1). Results Under the condition of low temperature and high speed (4 ℃,12 000×g), the identification effect was the best and the individual variation of the normal urine proteome was 0.635. Conclusions After acetone precipitation, the extraction effect of urine protein at low temperature and high speed (4 ℃, 12 000×g) is good. The individual variation of urine protein is similar to that of plasma proteome, which can be used for large-scale clinical analysis because of stability.

Key words: centrifugal temperature and speed, urine proteomics, individual variation

中图分类号:

R334

李京, 周冬冬, 孙伟. 离心条件对丙酮沉淀法提取尿蛋白质的影响[J]. 基础医学与临床, 2020, 40(2): 229-234.

LI Jing, ZHOU Dong-dong, SUN Wei. Effect of centrifugal conditions on urine protein extraction by acetone precipitation method[J]. Basic & Clinical Medicine, 2020, 40(2): 229-234.

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