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首页> 外文期刊>Science China Life Sciences >Comparison of alternative extraction methods for secretome profiling in human hepatocellular carcinoma cells
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Comparison of alternative extraction methods for secretome profiling in human hepatocellular carcinoma cells

机译：人肝癌细胞分泌蛋白谱分析的其他提取方法的比较

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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.
机译：分泌的蛋白质是早期发现和诊断疾病的重要来源，因此受到了极大的关注。从大量培养基中提取低浓度蛋白质，这些培养基富含盐和其他会干扰大多数蛋白质组学技术的化合物，这是分泌组研究的一个难题。超滤，沉淀和透析是可用于克服此问题的三种主要提取方法。本研究首次在没有偏倚的情况下比较了这三种方法的优缺点。离心超滤提供了最佳的提取效率，沉淀提供了最多的可识别蛋白质。这三种方法产生了与分泌组密切相关但又不同的信息。因此，应将它们视为补充方法或至少补充方法。鉴定出了360种独特的蛋白质，包括211种潜在的分泌蛋白质。与以前的研究相比，该研究还鉴定出42种新的分泌蛋白。本研究不仅为分泌酶组提取方法的选择提供参考，而且为肝细胞癌的研究扩展了分泌组数据库。

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