Antibodies with broad specificity to azaspiracids by use of synthetic haptens
Antibodies with broad specificity to azaspiracids by use of synthetic haptens
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DOI：
10.1021/ja066971h
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发表时间：
2006-11-29
影响因子：
15
通讯作者：
Miles, Christopher O.
中科院分区：
文献类型：
--
作者：
Forsyth, Craig J.;Xu, Jianyan;Miles, Christopher O.
目前迫切需要发展一种通用、灵敏、便携、定量的海洋毒素azaspiracid（AZA）检测方法。报道了使用含有AZA的环F-I的合成半抗原来产生通过其共同的C28-C40结构域与AZA交叉反应的抗体，以及这些抗体在ELISA和免疫亲和柱中的用途。这种方法具有许多优势，使用完整的azaspiracids（AZA）来自环境样品或全合成作为半抗原抗体的发展。左旋C28−C40氮杂螺酸结构域（1）的衍生物可以通过一锅法施陶丁格还原/分子内氮杂-维蒂希反应−亚胺捕获序列有效合成，以形成H-I环螺缩醛胺，并通过双分子内杂迈克尔加成组装成F-G环缩酮。将半抗原1与cBSA结合并在绵羊中免疫产生抗体，该抗体在不存在AZA 1的情况下识别并结合卵清蛋白结合的1。这种结合被1以浓度依赖性方式抑制。AZA 1、AZA 2、AZA 3和AZA 6的混合物引起与其总AZA含量一致的抗体结合抑制程度，而不仅仅是其AZA 1含量。该结果表明，抗体对AZA 2、AZA 3和AZA 6也具有与它们对AZA 1相似的亲和力，并且这些抗体适合于分析贝类样品中的AZA。
The development of general, sensitive, portable, and quantitative assays for the azaspiracid (AZA) class of marine toxins is urgently needed. Use of a synthetic hapten containing rings F−I of AZA to generate antibodies that cross-react with the AZAs via their common C28−C40 domain and use of these antibodies in ELISA and immunoaffinity columns are reported. This approach has many advantages over using intact azaspiracids (AZAs) derived from environmental samples or total synthesis as haptens for antibody development. A derivative of the levorotatory C28−C40 azaspiracid domain (1) was synthesized efficiently using a one-pot Staudinger reduction/intramolecular aza-Wittig reaction−imine capture sequence to form the H−I ring spiroaminal and a double intramolecluar hetero-Michael addition to assemble the F−G ring ketal. Conjugation of the hapten1to cBSA and immunization in sheep generated antibodies that recognized and bound to ovalbumin-conjugated1in the absence of AZA1. This binding was inhibited by1in a concentration-dependent manner. A mixture of AZA1, AZA2, AZA3, and AZA6 caused a degree of inhibition of antibody binding consistent with its total AZA content, rather than just its content of AZA1. This result suggests that the antibodies also have a similar affinity for AZA2, AZA3, and AZA6 as they do for AZA1 and that such antibodies are suitable for analysis of AZAs in shellfish samples.