L. Staunton , C. Tonry , R. Lis , S. Finn , J. O⿿Leary , M. Loda , M. Bowden , S.R. Pennington
{"title":"利用激光捕获显微解剖结合LC、MS、A技术报告分析前列腺癌肿瘤微环境蛋白质组","authors":"L. Staunton , C. Tonry , R. Lis , S. Finn , J. O⿿Leary , M. Loda , M. Bowden , S.R. Pennington","doi":"10.1016/j.euprot.2015.11.001","DOIUrl":null,"url":null,"abstract":"<div><p>Laser capture microdissection (LCM) allows microscopic procurement of specific cell types from tissue sections. Here, we present an optimized workflow for coupling LCM to LC⿿MS/MS including: sectioning of tissue, a standard LCM workflow, protein digestion and advanced LC⿿MS/MS. Soluble proteins extracted from benign epithelial cells, their associated stroma, tumor epithelial cells and their associated stromal cells from a single patient tissue sample were digested and profiled using advanced LC⿿MS/MS. The correlation between technical replicates was R<sup>2</sup> <!-->=<!--> <!-->0.99 with a mean % CV of 9.55%<!--> <!-->±<!--> <!-->8.73. The correlation between sample replicates was R<sup>2</sup> <!-->=<!--> <!-->0.97 with a mean % CV of 13.83%<!--> <!-->±<!--> <!-->10.17. This represents a robust, systematic approach for profiling of the tumor microenvironment using LCM coupled to label-free LC⿿MS/MS.</p></div>","PeriodicalId":38260,"journal":{"name":"EuPA Open Proteomics","volume":"10 ","pages":"Pages 19-23"},"PeriodicalIF":0.0000,"publicationDate":"2016-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.euprot.2015.11.001","citationCount":"12","resultStr":"{\"title\":\"Profiling the tumor microenvironment proteome in prostate cancer using laser capture microdissection coupled to LC⿿MS⿿A technical report\",\"authors\":\"L. Staunton , C. Tonry , R. Lis , S. Finn , J. O⿿Leary , M. Loda , M. Bowden , S.R. Pennington\",\"doi\":\"10.1016/j.euprot.2015.11.001\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Laser capture microdissection (LCM) allows microscopic procurement of specific cell types from tissue sections. Here, we present an optimized workflow for coupling LCM to LC⿿MS/MS including: sectioning of tissue, a standard LCM workflow, protein digestion and advanced LC⿿MS/MS. Soluble proteins extracted from benign epithelial cells, their associated stroma, tumor epithelial cells and their associated stromal cells from a single patient tissue sample were digested and profiled using advanced LC⿿MS/MS. The correlation between technical replicates was R<sup>2</sup> <!-->=<!--> <!-->0.99 with a mean % CV of 9.55%<!--> <!-->±<!--> <!-->8.73. The correlation between sample replicates was R<sup>2</sup> <!-->=<!--> <!-->0.97 with a mean % CV of 13.83%<!--> <!-->±<!--> <!-->10.17. This represents a robust, systematic approach for profiling of the tumor microenvironment using LCM coupled to label-free LC⿿MS/MS.</p></div>\",\"PeriodicalId\":38260,\"journal\":{\"name\":\"EuPA Open Proteomics\",\"volume\":\"10 \",\"pages\":\"Pages 19-23\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2016-03-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/j.euprot.2015.11.001\",\"citationCount\":\"12\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"EuPA Open Proteomics\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2212968515300246\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"EuPA Open Proteomics","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2212968515300246","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
Profiling the tumor microenvironment proteome in prostate cancer using laser capture microdissection coupled to LC⿿MS⿿A technical report
Laser capture microdissection (LCM) allows microscopic procurement of specific cell types from tissue sections. Here, we present an optimized workflow for coupling LCM to LC⿿MS/MS including: sectioning of tissue, a standard LCM workflow, protein digestion and advanced LC⿿MS/MS. Soluble proteins extracted from benign epithelial cells, their associated stroma, tumor epithelial cells and their associated stromal cells from a single patient tissue sample were digested and profiled using advanced LC⿿MS/MS. The correlation between technical replicates was R2 = 0.99 with a mean % CV of 9.55% ± 8.73. The correlation between sample replicates was R2 = 0.97 with a mean % CV of 13.83% ± 10.17. This represents a robust, systematic approach for profiling of the tumor microenvironment using LCM coupled to label-free LC⿿MS/MS.
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