Nanozeolite-driven approach for enrichment of secretory proteins in human hepatocellular carcinoma cells

Given the importance of secreted proteins as a source for early detection and diagnosis of disease, secreted proteins have been arousing considerable attention. However, the analysis of secreted proteins represents a challenge for cur-rent proteomic techniques. One of the difficulties in secretomic study is to concentrate proteins from large volume of growth media, particularly, the low abundant and low molecular weight proteins (molecular weight < 30 kDa). Herein, we describe a novel strategy for harvesting secretory proteins. In this approach, proteins secreted from the human hepatocellular carcinoma cell line were enriched by zeolite LTL nanocrystals, followed by 1-D SDS-PAGE for protein fractionation and then by LC-ESI-MS/MS for protein identification. In total, 1474 unique proteins were confidently identified, including 505 low molecular weight proteins, and covered a broad range of pI and molecular weight. Furthermore, this study not only offered an efficient and powerful method for the enrichment of secretory proteins but also allowed in-depth study of secretome of hepatocellular carcinoma cells. The reported work is expected to represent one of the most comprehensive secretomic analyses so far.