Currently, there are no reliable biomarkers of blood plasma for early detection of prostate cancer (PC), a one of the most common malignancies in men. This study developed, universalized and tested a new standardized methodology of detection of prostate cancer biomarkers--profiling of low-molecular weight proteome of blood plasma (1-17 kDa). This approach includes three main components: a preliminary sample preparation, time-of-flight mass spectrometry with an matrix-assisted laser desorption/ionization ALDI-TOF-MS), and the processing of data using bioinformatics software package. The potentials and prospects of the approach developed for identification of potential PC markers are demonstrated. 46 samples of blood plasma of PC patients and 26 controls were screened. This evaluation identified peptides/polypeptides that have the potential to be used for the detection of disease.