Abstract: Agarose microspheres (6 g/100 mL) were prepared by reversed-phase suspension regeneration method and they were cross-linked twice with epichlorohydrin and then coupled with the dye Cibacron Blue F3GA as a special ligand to form novel active microspheres (Blue Beads 6FF). The properties of the dye affinity medium were characterized by Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), pressure against flow velocity curve, swelling rate, leakage rate, and porosity. Our results showed that the microspheres had a good sphericity and uniform size with high porosity and swelling rate, a wide range of pH stability and strong pressure resistance. The average particle size of the microspheres was approximately 56.37 μm. The adsorption properties of the dye affinity medium were evaluated for lectin purification from black kidney beans. By fitting the typical Langmuir adsorption isotherm, the equilibrium adsorption capacity of lectin from black kidney beans was determined as 19.7 mg/mL microspheres. The dye affinity microspheres enabled rapid isolation and purification of lectin.

Key words: agarose microspheres, reversed-phase levitation regeneration method, Cibacron Blue F3GA, affinity chromatography, lectin from black kidney beans