College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, Hubei Province, People's Republic of China.
Appl Biochem Biotechnol. 2011 Jul;164(6):831-40. doi: 10.1007/s12010-011-9177-3. Epub 2011 Feb 1.
In this paper, a method of simultaneous immunizing BALB/c mice with staphylococcal enterotoxin (SE) A and B (SEA and SEB) to prepare a monoclonal antibody (3F2) for detecting both of SEA and SEB was developed. The results showed that antibody 3F2 had high titers against both SEA and SEB by enzyme-linked immunosorbent assay (ELISA). The sensitivities of 3F2 to SEA and SEB detected by ELISA were 133.2 and 82.5 ng/mL, respectively, and the detection limits for the two enterotoxins were about 1 ng/mL. The antibody 3F2 had high specificities and affinities to both SEA and SEB, and had no cross-reaction with SEC(1), bovine serum albumin, and ovalbumin. SEs-free skimmed milk samples were spiked with different concentrations of SEA, SEB, or both of them, respectively. Average recoveries of SEA and SEB from the spiked samples were all nearly between 82% and 104%. The result suggested that one cell fusion with simultaneous immunization by multiple antigen to prepare monoclonal antibody against them was possible, simple, and economic. The monoclonal antibody could be used in simultaneous detecting multifarious SEs.
本文介绍了一种同时免疫 BALB/c 小鼠以制备针对葡萄球菌肠毒素 A 和 B(SEA 和 SEB)的单克隆抗体(3F2)的方法,用于检测这两种肠毒素。酶联免疫吸附试验(ELISA)结果表明,抗体 3F2 对 SEA 和 SEB 具有高滴度。ELISA 检测 SEA 和 SEB 的灵敏度分别为 133.2 和 82.5 ng/mL,两种肠毒素的检测限约为 1 ng/mL。抗体 3F2 对 SEA 和 SEB 具有高特异性和亲和力,与 SEC(1)、牛血清白蛋白和卵清蛋白无交叉反应。在无 SE 的脱脂奶样品中分别加入不同浓度的 SEA、SEB 或两者的混合物。SEA 和 SEB 从加标样品中的平均回收率均在 82%至 104%之间。结果表明,通过多抗原同时免疫进行细胞融合以制备针对多种 SE 的单克隆抗体是可行的、简单的且经济的。该单克隆抗体可用于同时检测多种 SE。