一种家用收集和保存尿液蛋白质的方法  

A method for collecting and preserving urine protein in household

作　　者：秦伟伟 王婷 黄鹤 高友鹤 QIN WeiWei;WANG Ting;HUANG He;GAO YouHe(Beijing Key Laboratory of Genetic Engineering Drugs and Biotechnology, College of Life Sciences, Beijing Normal University, Beijing 100875, China)

机构地区：[1]北京师范大学生命科学学院基因工程药物及生物技术北京市重点实验室,北京100875

出　　处：《中国科学：生命科学》2018年第5期556-561,共6页Scientia Sinica(Vitae)

基　　金：国家重点研发计划(批准号:2016YFC1306300);国家重点基础研究发展计划(批准号:2013CB530805);北京自然科学基金(批准号:7173264;7172076);中央高校基本科研业务费专项资金资助(批准号:2015KJJCB21);北京合作建设项目(批准号:110651103);北京师范大学项目(批准号:11100704)资助

摘　　要：尿液是研究疾病早期标志物的重要来源.大规模收集尿液样本,建立尿液生物样本库具有长远的科学意义.本实验室曾提出用膜来吸附尿液蛋白质并干燥真空保存的方法.该方法具有经济便捷、占用存储空间小、样本可室温保存等特点,但其所需设备以及操作,仅适应于实验室.为方便全国各地的患者在家自行收集、保存尿液,并将样本室温运输,在此提出一种滤器联合注射器的简易装置.注射器提供压力,使尿液依次通过滤器内两层膜.第一层聚偏二氟乙烯膜隔离尿液中的细胞及碎片,第二层硝酸纤维素膜吸附尿液中蛋白质.结果显示,聚偏二氟乙烯膜有效地去除了尿液中细胞及碎片,具有很好的技术重复性.样本可室温保存1 0天,使样本远距离运输更加经济方便.Urine is a good source for identifying novel biomarkers.Urine samples should be comprehensively collected and preserved.Therefore, we established an urimem method in which urinary proteins were adsorbed onto membranes,following which the membranes were dried and stored in a vacuum bag.This method is economical, and samples can be stored at room temperature.In addition, it is more suitable in laboratories.For collecting urine samples of patients at home, a simple, easy to use method is necessary.The kit used in this method comprises a filter coupled with a syringe.The syringe was used as a pressure generator to pass the urine through the filter, which contained two membranes.The polyvinylidene fluoride （PVDF） membrane was used for isolating cells, and the nitrocellulose （NC） membrane was used for adsorbing proteins.After adsorption, the NC membrane was placed in a sealed plastic bag with a drying agent and stored at room temperature for 10 days.To assess the stability of the urimem method, SDS-PAGE and LC-MS/MS were performed to qualitatively and quantitative analyze the proteins that were eluted from the membrane.The results showed that the method was reproducible.This method is highly accessible and convenient for patients to collect urine by themselves at home.Furthermore, it facilitates sample transportation and may be beneficial for identifying research biomarkers in urine.

关 键 词：尿液 家用收集 室温保存 生物样本库 

分 类 号：R446.12[医药卫生—诊断学]