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用于免疫化学分析的基于聚合物分散体的免疫试剂。

Immunoreagents based on polymer dispersions for immunochemical assays.

作者信息

Pavlova I S, Generalova A N, Zubov V P, Zhorov O V, Martsev S P

机构信息

Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia.

出版信息

J Mol Recognit. 1998 Winter;11(1-6):185-7. doi: 10.1002/(SICI)1099-1352(199812)11:1/6<185::AID-JMR419>3.0.CO;2-7.

Abstract

Immunoreagents based on polymer dispersions consisting of unimodal polyacrolein (PAL) microspheres with diameters in the range 0.3-2.0 microns have been prepared and evaluated by various immunoassay techniques such as immunoradiometric assay of ferritin and microtitre particle agglutination and immunofiltration dot assay of group-specific polysaccharide of S. pyogenes (A-PS) in comparison with conventional carriers and methods. The antibodies were covalently or indirectly bound to the PAL. The coupled antibodies to ferritin retained a high average affinity (Ka = 4.5 x 10(9) M-1). In comparison with microcrystalline cellulose-based immunosorbent, more than an order-of-magnitude lower amount of PAL-IgG was necessary for the analysis of ferritin. Use of PAL-IgG gave a higher sensitivity of assay with a detection limit of 0.7 x 10(-13) M l-1 and a wider concentration range of antigen detection (about four orders of magnitude) without manifestation of the high-dose hook effect. Particle agglutination assay of A-PS in microtitre plate was shown to be a simple, demonstrative and highly sensitive one-step analytical method with a detection limit of 0.05 ng A-PS/ml or 10(4) cells/ml. The sensitivity of immunofiltration assay using both enzyme and latex markers was shown to be approximately the same (50 ng A-PS/ml) and the duration of the assay was 3-5 min. No cross-reaction of latex conjugates with non-A Streptococcus cell lysates were observed.

摘要

基于由直径在0.3 - 2.0微米范围内的单峰聚丙烯醛(PAL)微球组成的聚合物分散体的免疫试剂已被制备,并通过各种免疫测定技术进行评估,如铁蛋白的免疫放射测定、微量滴定板颗粒凝集以及化脓性链球菌(A - PS)群特异性多糖的免疫过滤斑点测定,并与传统载体和方法进行比较。抗体通过共价或间接方式与PAL结合。与铁蛋白偶联的抗体保留了较高的平均亲和力(Ka = 4.5 x 10(9) M-1)。与基于微晶纤维素的免疫吸附剂相比,分析铁蛋白时所需的PAL - IgG量降低了一个多数量级。使用PAL - IgG进行测定具有更高的灵敏度,检测限为0.7 x 10(-13) M l-1,抗原检测的浓度范围更宽(约四个数量级),且未出现高剂量钩效应。微量滴定板中A - PS的颗粒凝集测定被证明是一种简单、直观且高度灵敏的一步分析法,检测限为0.05 ng A - PS/ml或10(4) 个细胞/ml。使用酶和乳胶标记物的免疫过滤测定的灵敏度显示大致相同(50 ng A - PS/ml),测定持续时间为3 - 5分钟。未观察到乳胶缀合物与非A群链球菌细胞裂解物的交叉反应。

相似文献

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Immunoreagents based on polymer dispersions for immunochemical assays.用于免疫化学分析的基于聚合物分散体的免疫试剂。
J Mol Recognit. 1998 Winter;11(1-6):185-7. doi: 10.1002/(SICI)1099-1352(199812)11:1/6<185::AID-JMR419>3.0.CO;2-7.
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Polyacrolein microspheres as immunoreagents.
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