Purification of human serum immunoglobulins by Thiophilic chromatography (T-gel)

Abstract

Background and Objective: Human-derived antibodies, which are prepared by expensive methods, are used in diagnosis, prophylaxis, and treatment of diseases. The aim of this study was to develop an effective and inexpensive method for purification of human serum immunoglobulins. Methods: Human serum immunoglobulins were precipitated by ammonium sulfate (50% saturation) and then purification was done by Thiophilic chromatography (T-gel). Purification of antibodies was evaluated by cellulose acetate electrophoresis. Concentration of antibodies (IgG, IgM, IgA) in serum and the resulting pooled fractions were estimated by single radial immunodiffusion (SRID) and percentage of recovery was calculated. Findings: Analysis of products by electrophoresis showed only one band in γ-region. Recovery of the purified IgG was 75% while that for IgM was considerably low (about 17%). Conclusion: The purified immunoglobulins were eluated by a low salt buffer closely neutral pH, thus post treatment of product was not necessary. Thiophilic chromatography causes the purification of immunoglobulins at a stage after ammonium sulfate precipitation. The purified immunoglobulins in this study could be suitable for immunochemistry methods.

Keywords

• purification
• immunoglobulins
• human serum
• thiophilic gel