Comparison of alternative extraction methods for secretome profiling in human hepatocellular carcinoma cellsReport as inadecuate




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Science China Life Sciences

, Volume 54, Issue 1, pp 34–38

First Online: 21 January 2011Received: 11 June 2010Accepted: 30 August 2010

Abstract

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.

Keywordshepatocellular carcinoma secretome protein extraction ultrafiltration precipitation dialysis LC-MS-MS This article is published with open access at Springerlink.com

Electronic Supplementary MaterialSupplementary material is available for this article at 10.1007-s11427-010-4122-1 and is accessible for authorized users.

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Author: Jing Cao - ChengPin Shen - Jun Zhang - Jun Yao - HuaLi Shen - YinKun Liu - HaoJie Lu - PengYuan Yang

Source: https://link.springer.com/article/10.1007/s11427-010-4122-1







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