Kawatsu K, Hamano Y, Noguchi T
Department of Food Microbiology, Osaka Prefectural Institute of Public Health, Japan.
Toxicon. 1999 Nov;37(11):1579-89. doi: 10.1016/s0041-0101(99)00106-3.
For production of monoclonal antibodies against domoic acid, a causative agent of amnesic shellfish poisoning, three immunogens, domoic acid conjugated with bovine serum albumin (BSA), ovalbumin (OVA) and human gamma globulin (HGG), were prepared. The antiserum obtained from BALB/c mice immunized with domoic acid-BSA showed the highest affinity for domoic acid. The monoclonal antibody, DA-3, obtained from the mice was highly specific for domoic acid and showed a minor cross-reactivity with the isomers of domoic acid (isodomoic acids B, E, F, G and H), except for isodomoic acid A. Using DA-3 antibody, an indirect competitive enzyme immunoassay (idc-EIA) was developed for measurement of domoic acid. The working range for quantitative measurement of domoic acid and the quantification limit for domoic acid in shellfish were estimated to be 0.15-10 ng/ml and less than 0.04 microg/g, respectively. The mean recovery of domoic acid added to extracts of shellfish at toxin levels of 0.02 to 0.2 microg/ml was 103% with a coefficient of variation of 4.5%. The newly developed idc-EIA seems to be a useful method for monitoring domoic acid in shellfish.
为制备抗遗忘性贝类中毒病原体软骨藻酸的单克隆抗体，制备了三种免疫原，即与牛血清白蛋白（BSA）、卵清蛋白（OVA）和人γ球蛋白（HGG）偶联的软骨藻酸。用软骨藻酸-BSA免疫BALB/c小鼠获得的抗血清对软骨藻酸表现出最高亲和力。从小鼠获得的单克隆抗体DA-3对软骨藻酸具有高度特异性，除了软骨藻酸A外，与软骨藻酸的异构体（异软骨藻酸B、E、F、G和H）表现出轻微的交叉反应。使用DA-3抗体，开发了一种间接竞争酶免疫分析（idc-EIA）用于测定软骨藻酸。软骨藻酸定量测定的工作范围和贝类中软骨藻酸的定量限分别估计为0.15-10 ng/ml和小于0.04 μg/g。在毒素水平为0.02至0.2 μg/ml的情况下，添加到贝类提取物中的软骨藻酸的平均回收率为103%，变异系数为4.5%。新开发的idc-EIA似乎是监测贝类中软骨藻酸的一种有用方法。
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