Proteomics Analysis of Nasopharyngeal Carcinoma Cell Secretome Using a Hollow Fiber Culture System and Mass Spectrometry

• DOI: 10.1021/pr8006733
• Publication Date: 2008-11-14T19:01:50Z
• Authors: Hsin-Yi Wu; Ying-Hwa Chang; Yu-Chen Chang; Pao-Chi Liao
• Keywords: Secretory protein; Proteome; Lysis buffer

Secreted proteins, referred to as the secretome, are known regulate a variety of biological functions and involved in multitude pathological processes. However, some secreted proteins from cell cultures difficult detect because their intrinsic low abundance. They frequently masked by shed lysed cells substantial amounts serum used culture medium. We have proposed an analytical platform for sensitive detection utilizing hollow fiber (HFC) system coupled with proteomic approaches. The HFC enables high-density small volume where can be accumulated. In addition, lysis rates greatly reduced, which alleviates contamination cells. this study, nasopharyngeal carcinoma (NPC) were utilized evaluate efficiency collection analysis secretome. Cells adapted serum-free medium inoculated into system. rate was estimated 0.001−0.022%, determined probing four intracellular conditioned (CM), while 0.32−1.84% observed dish cultures. Proteins CM analyzed using SDS-PAGE liquid chromatography tandem mass spectrometry (LC-MS/MS). A total 134 identified 62 gel bands, 61% possess signal peptide and/or transmembrane domain. 37% secretome classified extracellular or membrane whereas 98% lysate proteins. suggest that may collect efficiently facilitate comprehensive characterization