用同系抗独特型单克隆抗体亲和层析法提纯小鼠单克隆抗体

Purification of Murine Monoclonal Antibody Using Affinity Chromatography of Syngenic Anti-idiotypic Monoclonal Antibody

1C_6McAb_2是抗7A_4McAb_1独特型的同系小鼠单克隆抗体。应用硫酸铵盐析,1C_6McAb_2—Sepharose 4B柱亲和层析法提纯了小鼠腹水7A_4McAb_1。获得的样品经二巯基乙醇还原,在SDS—PAGE上为两条带,分别是IgG的重链和轻链,免疫双扩散检查只与兔抗鼠IgG_1血清有沉淀反应,免疫电泳检查与兔抗鼠全血清反应只形成一条沉淀弧,纯化的McAb的PHA效价达125000/mg。本法提纯的McAb具有产量高,纯度好,特异性强等优点。

1C_6 McAb_2 is a syngenic murine monoelonal anti-body to the idiotype of 7A_4, monoclon al antibody (McAb_1). 7A_4 McAb_1, were extracted and purified from murine ascites by (NH_4)_2 SO_4 pre cipitation and 1C_6 McAb_2-Sepharose 4B affinity chromatography. Purified protein was reduced by 2 -mercaptoethanol was identified by SDS-PAGE. The result showed two bands which were heavy chain and light chain of IgG respectively. It was observed that a specific precipitation line with sera of rabbit against mice IgG and a single precipitation are with sera of rabbit against mice sera when double diffusion and immunoelectrophoresis were performed. The PHA titre of purified McAb reached 125 000/mg. The McAb purified by this method resulted in high yield, good purity and strong specificity.