猪初乳中SIgA纯化及其单抗制备

Purification of SIgA from porcine colostrums and preparation of its monoclonal antibody

为制备猪SIgA的特异性单克隆抗体,该研究采用饱和硫酸铵盐析法从猪初乳中提取猪SIgA蛋白,分别经Sephadex G-200凝胶层析、DEAE52纤维柱离子层析及Protein A柱亲合层析纯化。以纯化的猪SIgA蛋白为抗原免疫BALB/c小鼠制备抗猪SIgA的单克隆抗体。结果显示:已获得6株特异的抗猪SIgA的单克隆抗体,分别命名为2D9E6、2D9E9、3E1G2、3E1H5、4H5B7和4H5B10,6株单抗均为IgG1,κ型。将其中1株2D9E6制备腹水,经纯化后ELISA效价可达1∶81 000以上。表明以猪SIgA天然蛋白为免疫原,成功制备了6株高特异性的单克隆抗体,可为猪黏膜免疫研究提供基础材料。

To prepare for monoclonal antibody against porcine secretory immunoglobulin A（SIgA）,porcine SIgA was isolated and purified from colostrums by a procedure of precipitation with saturated ammonium sulfate,chromatography with sephadex G-200,ion-exchange chromatography with DEAE52 and affinity chromatography with protein A resin.The BALB/c mice were immunized four times with the purified SIgA.Six strains of hybridoma cells against porcine SIgA protein,named 2D9E6,2D9E9,3E1G2,3E1H5,4H5B7 and 4H5B10,were obtained through parallel screening with ELISA and Western blot.All six monoclonal antibodies belonged to mouse IgG1,κ isotype.The monoclonal antibody strain 2D9E6 was purified from hydroperitoneum,and its ELISA titer was higher than 1∶81 000.