论文部分内容阅读
为制备高纯度p24 抗原,在变性条件下,将大肠杆菌表达的重组p24 蛋白,使用Ni-NTA 亲和层析法进行了纯化。对包涵体中的p24 蛋白进行层析纯化时,回收率略有提高,但蛋白纯度(81% )低于菌体裂解纯化洗脱物(94% )。菌体中及纯化、复性后的目的蛋白均能与抗HIV-1 p24 单克隆抗体发生特异性反应。用纯化的p24 蛋白免疫小鼠,4 周时小鼠血清抗p24 平均抗体效价达1∶400。试验结果表明,大肠杆菌表达的HIV-1 p24 全菌体裂解物纯化后,可用作HIV-1 检测试剂的原料。
To prepare high purity p24 antigen, E. coli expressed recombinant p24 protein was purified using Ni-NTA affinity chromatography under denaturing conditions. The chromatographic purification of p24 protein in the inclusion body showed a slight increase in recovery, but the protein purity (81%) was lower than that of the bacterial lysate (94%). The mycelium, purified and refolded target protein reacted specifically with anti-HIV-1 p24 monoclonal antibody. Mice immunized with purified p24 protein, serum anti-p24 4 weeks mouse average antibody titer of 1: 400. The test results show that E. coli expressed HIV-1 p24 whole cell lysate purified, can be used as HIV-1 test reagents.