National Institute for Nutrition and Food Safety, Chinese CDC, Beijing 100021, China.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2010 Nov;27(11):1589-97. doi: 10.1080/19440049.2010.504237.
A monoclonal antibody against tetrodotoxin (TTX) was produced from the hybridoma cell line T6D9, which was established by the fusion of Sp2/0 myeloma cells with spleen cells isolated from a Balb/c mouse immunized with the TTX-keyhole limpet hemocyanin (KLH) conjugate. This monoclonal antibody belongs to the IgG1 subclass; the affinity constant of the antibody is 2.4 × 10(-8) mol l(-1). The relative cross-reactivity of the antibody with TTX was 100%, but with saxitoxin, KLH and bovine serum albumin (BSA) it was less than 1%, respectively. The titre of the antibody in ascites was 6.4 × 10(6); the reference working concentration was 1:1.2 × 10(5). By using this monoclonal antibody, a competitive indirect enzyme-linked immunoabsorbant assay (ELISA) for the analysis of TTX was developed. The linear portion of the dose-response curve of TTX concentration was in range 5-500 ng ml(-1). The limit of detection was 5 ng ml(-1) according 10% inhibition with TTX to anti-TTX monoclonal antibody. The concentration of TTX inhibiting 50% of antibody binding was about 50 ng ml(-1). The recoveries from TTX spiked samples were 79.5-109.5%. In addition, the toxicity of some wild puffer fish specimens captured from south-east China and the Yangzi River in Jiangsu province was determined. The results indicate that the toxicity and toxin tissue distribution vary in different species of wild puffer fish.
抗河豚毒素（TTX）单克隆抗体由杂交瘤细胞系 T6D9 产生，该细胞系由 Sp2/0 骨髓瘤细胞与经 TTX-贻贝血红蛋白（KLH）缀合物免疫的 Balb/c 小鼠脾细胞融合而成。该单克隆抗体属于 IgG1 亚类；抗体的亲和常数为 2.4×10(-8)mol l(-1)。该抗体与 TTX 的相对交叉反应性为 100%，而与石房蛤毒素、KLH 和牛血清白蛋白（BSA）的交叉反应性分别小于 1%。腹水抗体的效价为 6.4×10(6)；参考工作浓度为 1:1.2×10(5)。利用该单克隆抗体，建立了河豚毒素的竞争间接酶联免疫吸附测定（ELISA）分析方法。TTX 浓度的剂量反应曲线线性部分在 5-500ngml(-1)范围内。根据 TTX 对抗 TTX 单克隆抗体的 10%抑制作用，检测限为 5ngml(-1)。抑制 50%抗体结合的 TTX 浓度约为 50ngml(-1)。从河豚毒素添加样品中的回收率为 79.5-109.5%。此外，还测定了来自中国东南部和江苏省长江的一些野生河豚鱼标本的毒性。结果表明，不同种类的野生河豚鱼的毒性和毒素组织分布存在差异。
