一种高亲和力抗伊马扎吡单克隆抗体的研制及其在适合现场使用的ELISA中的结构

L. K. Siew, J. Dessi, J. Spoors, L. Winger, L. Jennens, C. Self
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引用次数: 0

摘要

尽管已经描述了除草剂imazapyr特异性的多克隆抗体,并形成了免疫测定的基础(一度在商业上可用,但后来被撤回),但很明显,该分子具有有限的免疫原性。我们评估了各种半抗原蛋白偶联物在多重免疫小鼠宿主中诱导抗体结合imazapyr的能力,通过半抗原竞争抑制与异源载体偶联的半抗原的反应性来评估。从一个高度反应的个体,我们随后开发了一种单克隆抗体,显示出对这种农用化学品的高反应性。有了这种抗体,我们已经能够配置适合实验室或现场使用的格式的免疫测定,表现出高性能和便利性,使imazapyr的灵敏度在1 ng/ml范围内可以实现。虽然需要在现场条件下进一步测试这种抗伊马扎吡抗体,但在开发有用的免疫分析方法以在需要时评估伊马扎吡时,获得持续的免疫反应性免疫球蛋白来源可能很重要。©2002 Wiley期刊公司化学工程学报(英文版);2001;DOI 10.1002 / fact.10012
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Development of a high affinity monoclonal antibody against imazapyr and its configuration in a sensitive ELISA suitable for field use
Although polyclonal antibodies specific for the herbicide imazapyr have been described and have formed the basis of an immunoassay (at one time commercially available but since withdrawn), it is clear that this molecule has limited immunogenicity. We evaluated various hapten–protein conjugates for their capacity to elicit antibody binding to imazapyr from multiply immunized murine hosts, as assessed by hapten-competed inhibition of reactivity with hapten conjugated to a heterologous carrier. From one highly reactive individual, we subsequently developed a monoclonal antibody exhibiting a high reactivity to this agrochemical. With this antibody we have been able to configure immunoassays in formats suitable for either laboratory or field use that exhibit both high performance and convenience, such that sensitivities for imazapyr in the range of 1 ng/ml can be achieved. While further testing of this anti-imazapyr antibody in field conditions is required, the availability of a constant source of immunoreactive immunoglobulin may be important in the development of useful immunoanalytical approaches for the assessment of imazapyr at the point-of-need. © 2002 Wiley Periodicals, Inc. Field Analyt Chem Technol 5: 281–287, 2001; DOI 10.1002/fact.10012
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