硫酸铵浓度对疏水作用层析纯化重组乙肝表面抗原的影响

以表面抗原活性收率和纯度为考察指标,转化乙肝病毒preS2-S基因重组酵母株,经甘油培养基充分增殖,转移到甲醇培养基中诱导表达.破碎细胞收集细胞原液,离心15000 r·min-1除去细胞碎片,超滤浓缩.结果表明,硫酸铵0.12、0.16 mol·L-1时盐浓度低,HBsAg未结合到疏水介质上;硫酸铵0.24 mol·L-1时疏水性强, HBsAg不宜从介质上洗脱.表明0.20 mol·L-1硫酸铵更适合重组HBsAg纯化.

Effec of ammonium sulfate concentration on purification of recombi nant HBsAg by hydrophobic interaction chromatography

To enhance the purification efficiency of recombinant hepatitis B surface antigen(HBsAg).Taking the recovery rate of the surface antigen activity and purity as inspection indicators.We transformed the recombinant yeast strains of hepatitis B virus preS2-S gene and sufficient proliferation of glycerol medium,then transferred to the methanol medium to induce expression.Breaking the cell and collecting cell essence,and removing cell debris after continuous centrifugal 15 000 r.min-1 and ultrafiltration concentration,to study the influence of hydrophobic medium for recombinant HBsAg adsorption and purification under different salt concentrations.The results showed that the low salt concentration of ammonium sulphate was 0.12 and 0.16 mol.L-1,and HBsAg failed to bind to the hydrophobic medium;strong hydrophobicity of ammonium sulphate was 0.24 mol.L-1,and HBsAg should not be eluted from the medium.Conclusion 0.20 mol.L-1ammonium sulfate was more suitable for the purification of recombinant HBsAg.