Preparation of Uniform-Sized Agarose Beads for Affinity Chromatography Matrix

Uniform-sized agarose beads were prepared by emulsification technique in this study. Agarose (4% w/v) was dissolved in boiling water containing 0.9% sodium chloride and used as water phase. A mixture of liquid paraffin containing 3 wt% of tween 80 emulsifier was heated to 70-80oC and used as oil phase. Then the heated oil phase was poured into the water phase and the mixture was stirred with a homogenizer to form uniform W/O emulsion. Then the emulsion was cooled down to room temperature under gentle agitation to form gel beads. The effect of emulsifier on the uniformity of the beads was investigated. The beads with different size can be prepared by using different concentration of emulsifier and the best result was found to be 3% w/v tween 80. Under optimized condition, the coefficient variation (C.V.) showing the size distribution of the beads was under 17%. Beads with diameter from 40 to 80 μm were obtained in this study