Two different approaches for developing immunometric assays of haptens.
Two different approaches for developing immunometric assays of haptens.
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开发半抗原免疫测定的两种不同方法。
DOI：
10.1093/clinchem/42.9.1532
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发表时间：
1996
影响因子：
9.3
通讯作者：
P. Pradelles
中科院分区：
文献类型：
--
作者：
J. Grassi;C. Créminon;Y. Frobert;E. Etienne;E. Ezan;H. Volland;P. Pradelles
为了改善小半抗原的免疫测定，我们开发了两种不同的方法，以非竞争性的形式进行测量。我们首先通过选择两组特异性针对肽的C-和N-末端部分的抗体来设计用于小肽（8-11个氨基酸）的双位点免疫测定。在每种情况下，测定灵敏度大大提高了相应的竞争性测定。更有趣的是，所有这些新的免疫测定法都比竞争性测定法更具特异性。在第二种方法中，我们开发了一种新的程序，固相固定化表位免疫测定（SPIE-IA），其中单个单克隆抗体使用相同的表位捕获和示踪剂结合，半抗原共价交联固相蛋白。迄今为止，SPIE-IA已成功地应用于测定带有伯氨基的半抗原，包括P物质、甲状腺素、白三烯C4、内皮素和血管紧张素II。在每种情况下，测定灵敏度均显著提高。
To improve immunoassays of small haptens, we developed two different approaches for their measurement in a non-competitive format. We first devised two-site immunometric assays for small peptides (8-11 amino acids) by selecting two sets of antibodies specifically directed against C- and N-terminal moieties of the peptides. In each case, assay sensitivity improved substantially over that of the corresponding competitive assays. More interestingly, all of these new immunometric assays were much more specific than the competitive assays. In a second approach, we developed a new procedure, solid-phase-immobilized epitope immunoassay (SPIE-IA), in which a single monoclonal antibody uses the same epitope for capture and tracer binding and the hapten is covalently cross-linked to solid-phase proteins. To date, SPIE-IA have been successfully applied to the determination of haptens bearing primary amino groups, including substance P, thyroxine, leukotriene C4, endothelin, and angiotensin II. In each case, assay sensitivity was significantly improved.