Serum proteomics using mass spectrometry

The identification and eventual application of tumor markers in cancer screening, early detection, diagnosis, and prognosis is a continuing focus of significant translational cancer research. While many new candidate markers have been discovered and at least partly characterized, very few have found widespread clinical application limited presently to the use of CA-125 in ovarian cancer, CEA, primarily in colon cancer, and PSA in prostate cancer screening and patient monitoring. The rapidly emerging field of cancer genomics and proteomics, and their clinical translation as "molecular diagnosis" and "molecular medicine" are already beginning to transform the field, and the accelerating growth of information and technology in this research area will undoubtedly transform the field of tumor markers and their application in the near future leading to improved molecular tools for cancer diagnosis, prognosis, and treatment and ultimately, to the emergence of novel and more effective cancer therapies, including improved approaches for immunotherapy and cancer prevention strategies. Toward this goal, herein are described detailed methods and workflows for mass spectrometry-based biomarker discovery in serum/plasma utilizing two complementary approaches-matrix-assisted laser desorption ionization time of flight (MALDI-TOF) and nanoflow reversed-phase liquid chromatography (RPLC)-tandem mass spectrometry (MS/MS). These discovery workflows incorporate both abundant protein depletion and sample fractionation upstream of analytical mass spectrometry to optimize the identification and quantitation of lower abundant species.