摘要
采用不同的方法纯化相思子毒素单抗,寻求一种效果好、操作简便的纯化方法。采用硫酸铵沉淀法、辛酸-硫酸铵法和硫酸铵-蛋白A亲和层析法三种方法分离纯化相思子毒素单抗,并采用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)、BCA蛋白检测试剂盒(BCA Protein Assay Kit)和双抗夹心酶联免疫吸附试验(ELISA)对纯化产物进行相对分子质量、浓度、纯度、回收率及免疫活性的鉴定。结果表明,纯化后单抗的回收率以硫酸铵沉淀法及辛酸-硫酸铵法较高,硫酸铵-蛋白A亲和层析法较低;产物纯度以硫酸铵-蛋白A亲和层析法和辛酸-硫酸铵法较高,硫酸铵沉淀法较差;不同方法纯化后的单抗活性未见降低。
The experiments were conducted to select good methods for the purification of abrin monoclonal antibody. The methods used included ammonium sulfate precipitation,caprylic acid-ammonium method and protein A chromatography. The relative molecular weight, concentration, purity, recovery rate and immunological activity of parified products were examined by SDS-PAGE, BCA protein assay, ELISA. The results showed that the recovery rates of purified monoclonal antibody were higher by using ammonium sulfate precipitation and caprylic acid-ammonium method than that by using protein A chromatography. The purities of monoclonal antibody were higher by using protein A chromatography and caprylic acid-ammonium method than that by using ammonium sulfate precipition. The activities of monoclonal antibody purified with different methods were normal.
出处
《动物医学进展》
CSCD
2006年第11期72-75,共4页
Progress In Veterinary Medicine
基金
国家自然基金项目(30230260)
