A Docile Method of Isolation and Purification of Riboflavin Binding Protein (Rfbp) from Peahen ( Pavo cristatus ) and Domestic Fowl ( Gallus gallus )

Riboflavin binding protein (Rfbp) was isolated from domestic fowl (Gallus gallus) and peahen (Pavo cristatus) egg-white and egg-yolk. The protein was purified in two steps, DEAE-Sephadex A-50 ion exchange chromatography and eluted with phosphate buffer pH 7.3 containing 0.5 M sodium chloride. The final purification of protein was achieved on Sephadex G-100. The purity of the protein was judged on cylindrical and slab gel electrophoresis, SDS-PAGE technique. Sephadex G-100 fraction Rfbp moved as a single band both on the Slab and Cylindrical gels. Comparison of the mobility of Rfbp with that of the standard molecular weight marker proteins revealed with that the Rfbp had a molecular weight close to 29,000 kd. Interestingly, hen egg-white Rfbp and peahen egg- white, yolk Rfbp had the same molecular weight as revealed by the SDS-PAGE. This is a novel approach for the study of riboflavin binding protein purified from different avian eggs in two steps and studied electrophoretic characterization with standard molecular weight marker.