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针对通过N-甲基脱氢丙氨酸偶联的微囊藻毒素的单克隆和多克隆抗体的制备及其特异性

Production and specificity of mono and polyclonal antibodies against microcystins conjugated through N-methyldehydroalanine.

作者信息

Mikhailov A, Härmälä-Braskén A S, Meriluoto J, Sorokina Y, Dietrich D, Eriksson J E

机构信息

Department of Biology, Laboratory of Animal Physiology, University of Turku, FIN-20014, Turku, Finland.

出版信息

Toxicon. 2001 Apr;39(4):477-83. doi: 10.1016/s0041-0101(00)00148-3.

Abstract

Microcystins (MCs) are a group of closely related toxic cyclic heptapeptides produced by common cyanobacteria (blue-green algae). Their toxicity is associated with specific inhibition of intracellular protein phosphatases type-1 and type-2A (PP1 and PP2A, respectively). We have developed a battery of antibodies to microcystins using chemical modification (aminoethylation) of one of its core amino acids, N-methyl-dehydroalanine. The developed antibodies displayed different reactivities to closely related MCs. Selected monoclonal antibodies were used for quantitative competitive ELISA assays. The analytical sensitivity of these assays was up to 1 ng/ml. Comparison of the developed ELISA tests with HPLC-based measurements of MCs in laboratory and field samples showed a good correspondence between the results yielded by these two methods. The antibodies developed by this technique provide the means for developing extremely sensitive and specific analytical assays for direct measurement of toxins in cyanobacterial or water samples.

摘要

微囊藻毒素(MCs)是由常见蓝藻(蓝绿藻)产生的一组密切相关的有毒环状七肽。它们的毒性与细胞内1型和2A型蛋白磷酸酶(分别为PP1和PP2A)的特异性抑制有关。我们通过对其核心氨基酸之一N-甲基脱氢丙氨酸进行化学修饰(氨乙基化),开发了一系列针对微囊藻毒素的抗体。所开发的抗体对密切相关的微囊藻毒素表现出不同的反应性。选择的单克隆抗体用于定量竞争ELISA测定。这些测定的分析灵敏度高达1 ng/ml。将所开发的ELISA测试与实验室和现场样品中基于HPLC的微囊藻毒素测量结果进行比较,结果表明这两种方法所得结果具有良好的一致性。通过该技术开发的抗体为开发极其灵敏和特异的分析方法以直接测量蓝藻或水样中的毒素提供了手段。

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