Rapid and highly sensitive enzyme immunoassay for quantitative determination of tetrodotoxin
Rapid and highly sensitive enzyme immunoassay for quantitative determination of tetrodotoxin
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DOI：
10.7883/yoken1952.50.133
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发表时间：
1997-06-01
期刊：
影响因子：
--
通讯作者：
Shibata, T
中科院分区：
文献类型：
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作者：
Kawatsu, K;Hamano, Y;Shibata, T
用河豚毒素（TTX）-牛血清白蛋白（BSA）偶联物免疫Balb/c小鼠，获得抗TTX的单克隆抗体。该单克隆抗体对TTX具有高度特异性，与TTX衍生物河豚酸或gonyautoxins无交叉反应，但观察到与脱水河豚毒素有轻微交叉反应。该单克隆抗体可中和TTX的致死活性。利用该单克隆抗体，建立了快速、高灵敏度的TTX竞争酶免疫分析法。用竞争性酶免疫分析系统定量测定TTX的时间约为30 min（若包括固相抗原制备时间则需90 min），定量分析TTX的工作范围为2-100 ng/ml。在回收试验和TTX样品的检查中，小鼠生物测定和EIA分析的结果相关性良好（r = 0.987）。此外，它表明，低浓度的TTX，这不能检测到的小鼠生物测定，可以定量测定的竞争性EIA。
A monoclonal antibody against tetrodotoxin (TTX) was obtained from Balb/c mice immunized with TTX-bovine serum albumin (BSA) conjugate. The monoclonal antibody was highly specific for TTX and had no cross-reaction to tetrodonic acid, which is a TTX derivative, or gonyautoxins, although a minor cross-reaction to anhydro-tetrodotoxin was observed. The monoclonal antibody neutralized the lethal activity of TTX. By using the monoclonal antibody, a rapid and highly sensitive competitive enzyme immunoassay (EIA) for quantitative analysis of TTX was developed. By the competitive EIA system, TTX can be determined quantitatively in about 30 min (90 min are required if the time for preparation of the solid-phase antigen was included), and the working range for quantitative analysis of TTX was 2-100 ng/ml. In recovery tests and examinations of TTX samples, results of the mouse bioassay and EIA analyses correlated well (r = 0.987). Moreover, it was demonstrated that low concentrations of TTX, which could not be detected by the mouse bioassay, could be determined quantitatively by the competitive EIA.