Comparison of alternative extraction methods for secretome profiling in human hepatocellular carcinoma cells  

作　　者：CAO Jing SHEN ChengPin ZHANG Jun YAO Jun SHEN HuaLi LIU YinKun LU HaoJie YANG PengYuan 

机构地区：[1]Institutes of Biomedical Sciences and Department of Chemistry, Fudan University, Shanghai 200032, China [2]Liver Cancer Institute Zhongshan Hospital, Fudan University, Shanghai 200032, China

出　　处：《Science China(Life Sciences)》2011年第1期34-38,共5页中国科学（生命科学英文版）

基　　金：supported by the National Natural Science Foundation of China (Grant No. 209750240);the National Basic Research Program of China (Grant No. 2010CB912700)

摘　　要：Secreted proteins are important sources for early detection and diagnosis of disease, and as such have received considerable attention. The extraction of low concentration proteins from large volumes of culture media, which are rich in salts and other compounds that interfere with most proteomics techniques, presents a problem for secretome studies. Ultrafiltration, precipitation, and dialysis are three major extraction methods that can be used to overcome this problem. The present study for the first time, compared the merits and shortcomings of these three methods, without bias. Centrifugal ultrafiltration provided the best extraction efficiency, and precipitation provided the highest number of identifiable proteins. The three methods yielded closely related, but different, information on the secretome; thus, they should be considered complementary or, at least, supplementary methods. Three hundred and sixty unique proteins were identified, including 211 potential secreted proteins. Compared with previous studies, this study also identified 42 new secreted proteins. The present study not only offers a reference for the selection of secretome extraction methods, but also expands the secretome database for the investigation of hepatocellular carcinoma.Secreted proteins are important sources for early detection and diagnosis of disease, and as such have received considerable attention. The extraction of low concentration proteins from large volumes of culture media, which are rich in salts and other compounds that interfere with most proteomics techniques, presents a problem for secretome studies. Ultrafiltration, precipitation, and dialysis are three major extraction methods that can be used to overcome this problem. The present study for the first time, compared the merits and shortcomings of these three methods, without bias. Centrifugal ultrafiltration provided the best extraction efficiency, and precipitation provided the highest number of identifiable proteins. The three methods yielded closely related, but different, information on the secretome; thus, they should be considered complementary or, at least, supplementary methods. Three hundred and sixty unique proteins were identified, including 211 potential secreted proteins. Compared with previous studies, this study also identified 42 new secreted proteins. The present study not only offers a reference for the selection of secretome extraction methods, but also expands the secretome database for the investigation of hepatocellular carcinoma.

关 键 词：hepatocellular carcinoma SECRETOME protein extraction ULTRAFILTRATION precipitation DIALYSIS LC-MS/MS 

分 类 号：TQ655[化学工程—精细化工] S852.618[农业科学—基础兽医学]