纯化小鼠IgM腹水单克隆抗体方法的探索

Comparision of Purification Methods on IgM Monoclonal Antibodies from Mouse Ascitic Fluid

本文选用了混有大量IgG的小鼠IgM腹水单克隆抗体(Ps-1)。采取三种不同方案提纯抗体:A、盐析一层析法;B、低渗透析一层析法,即单克隆抗体(McAb)透析后所得的沉淀及上清部份分别层析;C、二次层析法。比较了它们的蛋白得率、免疫活性和纯度。A、B、C方案所得抗体蛋白量分别是0.62,1.43,2.2mg/ml腹水,各占腹水蛋白的3.8％,9.0％,14％。电泳纯度在90％左右,Ig亚型鉴定均是к链的IgM抗体,并与IgG、IgA无交叉。提纯后抗体均保持免疫活性。

This paper reported three programse of purification of IgM monoclonal antibody (PS-1) from mouse ascitic fluid, ie.( A) neutral salt(NH4)2SO4 precipitation,(B) dialysis against low ionic strength buffer, and (C) twice purification by gd filtration chromatography on Sephacryl S-200. The available protein rates,immunologic activities,and purities of production by each programme were compared Antiboby protein contorts of A, B, C were 0.62,1.43, and 2.2 mg/ml of ascitic fluid respectively. They were about 3.8%, 9.0% and 14% of asdtic fluid protein respectively. Sectrophoresis purities of all productions were about 90%. Immunologic activity of McAb produced by C programme was higher than that by A and R In addition, all of them have no cross reaction with IgG, IgA. The purified McAb can be stored for half year in 4C