Montagne P, el Omari R, Cliquet F, Cuillière M L, Duheille J
Immunology Laboratory, Faculty of Medicine, Vandoeuvre les Nancy, France.
Bioconjug Chem. 1992 Nov-Dec;3(6):504-9. doi: 10.1021/bc00018a007.
A new microparticle-enhanced nephelometric immunoassay has been recently described as a sensitive, accurate, and easy-to-perform competitive immunoassay for various analytes. As initially described, this test is based on the nephelometric quantification of the inhibition, by the antigen to be assayed, of immunoagglutination of microparticle-antigen conjugates. Its applicability as a competitive immunoassay is thus limited by the necessary availability of pure antigens to prepare microparticle-antigen conjugates. In this paper, we report an adaptation of this initial test, where microparticles are coated by monoclonal antibodies, eliminating the need for purified antigens. The new configurations of particle agglutination-based immunoassays described include use of these microparticle-antibody conjugates with microparticle-antigen conjugates, free antigen, and anti-mouse immunoglobulins antiserum. The feasibility of such configurations is studied with human chorionic gonadotropin hormone, human thyroid stimulating hormone, and human myoglobin as antigens. Capture of the analyte by microparticle-antibody conjugates is evidenced by inhibition of their agglutination with microparticle-antigen conjugates and by agglutination in a sandwich assay with a complementary monoclonal antibody or polyclonal antiserum. The use of a second xenogenic antibody enhances the agglutination process and increases the assay sensitivity. Microparticle-antibody conjugates may extend the applications of microparticle-enhanced nephelometric immunoassays to unavailable analytes.
最近,一种新型微粒增强散射比浊免疫测定法被描述为一种灵敏、准确且易于操作的针对各种分析物的竞争性免疫测定法。如最初所描述的,该测试基于待测定抗原对微粒 - 抗原缀合物免疫凝集的抑制作用的散射比浊定量。因此,其作为竞争性免疫测定法的适用性受到制备微粒 - 抗原缀合物所需纯抗原可用性的限制。在本文中,我们报告了对该初始测试的一种改进,其中微粒用单克隆抗体包被,从而无需纯化抗原。所描述基于颗粒凝集的免疫测定法的新配置包括这些微粒 - 抗体缀合物与微粒 - 抗原缀合物、游离抗原和抗小鼠免疫球蛋白抗血清的使用。以人绒毛膜促性腺激素、人促甲状腺激素和人肌红蛋白作为抗原研究了这种配置的可行性。微粒 - 抗体缀合物对分析物的捕获通过其与微粒 - 抗原缀合物凝集的抑制以及在与互补单克隆抗体或多克隆抗血清的夹心测定中的凝集来证明。使用第二种异种抗体可增强凝集过程并提高测定灵敏度。微粒 - 抗体缀合物可能会将微粒增强散射比浊免疫测定法的应用扩展到难以获得的分析物。
J Clin Lab Anal. 1992
J Clin Lab Anal. 1996