TY - JOUR
AU - Lubman, David M.
AB - A two‐dimensional (2‐D) liquid phase separation method, liquid isoelectric focusing followed by nonporous reversed‐phase high performance liquid chromatography (HPLC), was used to separate proteins from human ovarian epithelial whole cell lysates. HPLC eluent was interfaced on‐line to an electrospray ionization (ESI) time of flight (TOF) mass spectrometer to obtain accurate intact protein molecular weights (Mr). 2‐D protein expression maps were generated displaying protein isoelectric point (pI) versus intact protein Mr . Resulting 2‐D images effectively displayed quantitative differential protein expression in ovarian cancer cells versus non‐neoplastic ovarian epithelial cells. Protein peak fractions were collected from the HPLC eluent, enzymatically digested, and analyzed by matrix‐assisted laser desorption/ionization (MALDI) TOF‐mass spectrometry (MS) peptide mass fingerprinting and by MALDI‐quadrupole TOF tandem mass spectrometry peptide sequencing. Interlysate comparisons of differential protein expression between two ovarian adenocarcinoma cell lines, ES2 and MDAH‐2774, and ovarian surface epithelial cells was performed. Five pI fractions from each sample were selected for comparative study and over 300 unique proteins were positively identified from the 2‐D liquid expression maps using MS, which covered around 60% of proteins detected by on‐line ESI‐TOF‐MS. This represents one of the most comprehensive proteomic analyses of ovarian cancer samples to date. Protein bands with significant up‐ or down‐regulation in one cell line versus another as viewed in the 2‐D expression maps were identified. This strategy may prove useful in identifying novel ovarian cancer marker proteins.
TI - Comprehensive proteome analysis of ovarian cancers using liquid phase separation, mass mapping and tandem mass spectrometry: A strategy for identification of candidate cancer biomarkers
JF - Proteomics
DO - 10.1002/pmic.200300763
DA - 2004-08-01
UR - https://www.deepdyve.com/lp/wiley/comprehensive-proteome-analysis-of-ovarian-cancers-using-liquid-phase-GCOXk0dkvI
SP - 2476
EP - 2495
VL - 4
IS - 8
DP - DeepDyve
ER -