Biotine-based new affinity solid support enhancement for purification of avidin

In this study, a new affinity solid support was developed for the purification of Avidin protein from egg white. Within the scope of this study, a biotin-based cryogel column which exhibits specific affinity towards Avidin was developed in the presence of cross-linkers. The characterization of the synthesized cryogel column was carried out by swelling test, FT-IR and SEM analyses. In another step of the study, the Avidin was purified from the crude extract by using the synthesized cryogel separation column. The protein content of the purified Avidin was determined and the Avidin binding to the cryogel column, optimization studies and reusability tests were performed by UV spectrophotometric measurements. In addition, Circular Dichroism (CD) Spectroscopy was used to evaluate the changes in secondary and tertiary structures. The molecular weight of the purified Avidin was determined by Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE).