应用A蛋白亲和层析法纯化单克隆抗体

Purification of Monoclonal Antibodies Using Protein A Affinity Chromatography

应用Protein A亲和层析法,从采集的小鼠腹水中纯化出了抗凝血因子Ⅶ单克隆抗体,用SDS-PAGE和ELISA法分别检测了纯化后单克隆抗体的纯度及效价,结果显示,电泳为两条带,分别为免疫球蛋白G(IgG)的重链和轻链,纯化后的单克隆抗体纯度达到电泳纯,应用间接ELISA法测定腹水效价为1×10-7左右,与未纯化前无差异。结果表明,应用A蛋白亲和层析法能够得到纯度较高的单克隆抗体,适用于高纯度单克隆抗体的制备。

The antibodies agaist human blood clotting factor Ⅶ were purified by using affinity chromatography with protein A sepharose CL-4B. The titers and purity of the purified monoclonal anti-FⅦ were determined by indirect ELISA and SDS-PAGE respectively. The result of SDS-PAGE had reached the srandard of electrophoretic purity. The titers of the purified antibody in ascetic fluid were about 1 10^-7 by indirect ELISA. The results indicated that the antibodies of higher purity had been prepared by using protein A affinity chromatography, which were fit for the preparation highly purified monoclonal antibodies.