Rapid screening of protein profiles of human breast cancer cell lines using non-porous reversed-phase high performance liquid chromatography separation with matrix-assisted laser desorption/ionization time-of-flight mass spectral analysis

Non-porous reversed-phase (NP-RP) HPLC has been used to rapidly generate pr otein profiles of whole cell lysates of human breast cancer cell lines. The non-porous packing material used was silica coated with C18, which provide d rapid separation with high collection efficiency of proteins from cell ly sates. This method was used to study the differences in protein profiles am ong normal cells and fully malignant cells that share a common genetic back ground. The highly expressed proteins in each cell type were separated and collected in the liquid state where they were analyzed by matrix-assisted l aser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) t o obtain the molecular weight of tbe proteins. The protein fractions were t hen subjected to tryptic digestion and analyzed by pulsed delay extraction (PDE)-MALDI-TOFMS to obtain the peptide maps. The expressed proteins were i dentified based upon the molecular weight and peptide map using database-se arching procedures. It is shown that key cancer-related proteins can be det ected and identified which may be potentially used as biomarkers for cancer detection. Copyright (C) 1999 John Wiley & Sons, Ltd.