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试验为制备抗κ-酪蛋白单克隆抗体并对其进行特性鉴定,采用足垫免疫κ-酪蛋白的方法刺激BALB/c小鼠产生免疫应答,将小鼠腘淋巴结细胞与骨髓瘤细胞在PEG诱导下进行融合,通过克隆化培养获得了能稳定分泌抗κ-酪蛋白抗体的杂交瘤细胞株,并利用Protein G亲和层析柱纯化腹水获得了单克隆抗体。试验获得了3株杂交瘤细胞株:1C4、3G3、3E6,所分泌的抗体亚类均为IgG1,其中3G3与3E6抗原识别表位不同,1C4与3E6抗原识别表位相近,1C4株腹水纯化后效价为1.28×106,其亲和力常数为2.89×108 mol/L。试验制备出了亲和力较好的抗κ-酪蛋白单克隆抗体,为牛奶酪蛋白的快速检测奠定了基础。
In order to prepare and characterize the anti-κ-casein monoclonal antibody, BALB / c mice were immunized with kappa-casein by footpad. Immune response was induced in mice with lymph node cells and myeloma cells in PEG After cloning, the hybridoma cell line which can stably secrete anti-κ-casein antibody was obtained. The monoclonal antibody was obtained by purification of ascites by Protein G affinity chromatography. Three hybridoma cell lines, 1C4, 3G3 and 3E6, were obtained. The secreted antibody subclasses were all IgG1. Among them, 3G3 and 3E6 antigen epitopes were different. 1C4 and 3E6 antigen epitopes were similar. After 1C4 ascites fluid purification The titer was 1.28 × 106, and its affinity constant was 2.89 × 108 mol / L. Experiments to prepare a good affinity of anti-κ-casein monoclonal antibody for the rapid detection of milk casein laid the foundation.