Serum Removal - Proteomics of Serum samples

Comparative analysis of normal & infected human patients’ peripheral blood serum proteome yields a potential approach for diagnostic or therapeutic monitoring of disease prognosis. However, the presence of salts, lipids, and abundant globular & albumin-proteins mask identification of very small and actual biomarkers of the serum. In two-dimensional electrophoresis (2-DE) it is crucial to deplete the masking biomolecules thereby enriching the actual serum biomarker in order to critically analyze the serum proteome data. We had earlier used a wide variety of in-house or traditional laborious protocols to deplete unwanted masking biomolecules – like treatment of serum samples with SDS, ammonium sulfate precipitation, etc. which were not yielding expected results following Isoelectric focusing (IEF) of serum samples using IPG strips. Finally, to deplete the high abundant and masking biomolecules from human serum samples, we adopted a kit from SERVA Electrophoresis Company called “SERVA BluePrep Major Serum Protein Removal Kit”. As described in the kit protocol, we got clean super enriched human serum samples and the same was experimentally confirmed by SDS-PAGE (4-10% gel).