Improved two-dimensional gel electrophoresis representation of serum proteins by using ProtoClear (TM)

ProtoClear(TM) is a proprietary technique for clearing albumin and immunogl obulin G (IgG) from human serum samples. Albumin constitutes 57-71% of tota l serum protein and IgG ranges from 8-26%. Removal of these two proteins al one clears similar to 75% of the total protein present in serum and allows the detection of the remaining proteins that are present in far lower conce ntrations. ProtoClear effectively removed > 95% of human serum albumin (HSA ) and > 97% of human IgG as measured by an anti-HSA competitive immunoassay and a radial immunodiffusion assay, respectively. ProtoClear was far more specific at removing albumin and IgG than Cibracon Blue Dye chromatography (Cibracon Blue), the typically utilized alternative. Comparing two-dimensio nal (2-D) gels of serum cleared by either Cibracon Blue or by ProtoClear, i t was apparent that Cibracon Blue removed a number of proteins in addition to albumin. Following removal of albumin and IgG from serum, we found a sig nificant improvement in the resolution of polypeptide spots detected on two -dimensional gels.