Research On Bio-separation By Domestic Monodisperse Polystyrene Microspheres

Monodisperse polystyrene microsphere, with high rigidity and wide pH stability range, which is the deficiency of silica-based RPC matrices, for example ODS C₁₈, are efficient HPLC packing materials for separation process.The optical gradient of separation of IL-2 by domestic Microspheres RPC column packed with monodisperse polystyrene microspheres was that the mobile phase B (0.1%TFA+ 80%CH₃CN ) was linear increased from 0% to 100% in thirty minutes while the mobile phase A was 0.1%TFA. The domestic Microspheres RPC column was evaluated by the Systemic Test according to the Chinese pharmacopoeia on the optical gradient condition. The results showed that the column efficiency, resolution, reproducibility, tailing factor of domestic Microspheres RPC column were in good agreement with the requirement of Pharmacoloeia, and the column efficiency, resolution, backpressure and polarity of domestic Microspheres RPC column were equivalent to those of SOURCE column. The range of pH stability of domestic Microspheres RPC column was 1-14.And backpressure and non-polarity of domestic Microspheres RPC column were lower than those of Hypersil C₈ and Polaris C₁₈, which was in favor of the high flow-rate and high recovery of bioactivity. And the maximum amount of IL-2 on domestic Microspheres RPC column(l50×4.6mm) was 300μg. According to the scale factor, the flow-rate and amount load of domestic Microspheres RPC preparative column were 7.9mL/min and 2.36mg respectively. The purity and bioactivity of the separated IL-2 met the requirements of quality control.To develop the application of separation of bio-molecular by domestic Microspheres RPC column, in this paper, the separations of aescin were obtained by isocratic elution, while the protein(bovine serum albumin BSA and ovum albumin OVA) were isolated by line gradient elution. The results of isocratic elution on domestic Microspheres RPC column (MKF-C18) showed that 4 component of aescin were separated in baseline when the acetonitrile volume fraction was 32%, and there was a good line relationship between the lnk' of 4 component and the acetonitrile...