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  

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

作　　者：蔺芳[1] 

机构地区：[1]新乡学院生命科学与生物技术系,河南新乡453003

出　　处：《东北农业大学学报》2012年第9期135-138,共4页Journal of Northeast Agricultural University

基　　金：河南省政府决策研究课题(B578);2010年度河南教育学院青年科研课题(20100103)

摘　　要：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15 000 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。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.

关 键 词：疏水作用层析 纯化 重组乙肝表面抗原 硫酸铵 

分 类 号：G424.31[文化科学—课程与教学论]