摘要
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。
To obtain the anti kappa casein monoclonal antibody and complete the identification of the antibody characteris- tics. The BALB/c mice were immunized with kappa casein using foot-pad immunization. Popliteal lymph node cells from the immunized mice were fused with SP2/0 myeloma cells in the presence of PEG. Three hybridoma strains （1C4, 3G3, 3E6） which secreted the antibody specific for kappa casein were obtained. The sub-class of the antibodies were IgG1. The ascites were purified by Protein G affinity layer absorption column. The antigenic epitope of 3G3 and 3E6 were different and it was close between 1C4 and 3E6. The titer of purified ascites（1C4） was 1.28 × 10^6 and the affinity constant was 2.89 × 108 mol/L. A anti-kappa casein monoclonal antibody with good affinity had been achieved,which provided foundations for the rapid detection of casein in bovine milk samples.
出处
《中国畜牧兽医》
CAS
北大核心
2014年第3期24-27,共4页
China Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(61171022、60971011)
关键词
κ-酪蛋白
单克隆抗体
特性鉴定
kappa casein
monoclonal antibody
characteristics identification
