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使用合成肽Cys-env gp46(188 - 224)作为抗原,对血清中(抗人I型T细胞白血病病毒)IgG进行灵敏的酶免疫测定(免疫复合物转移酶免疫测定)。

Sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for (anti-human T-cell leukemia virus type I) IgG in serum using a synthetic peptide, Cys-env gp46(188-224), as antigen.

作者信息

Kohno T, Sakoda I, Ishikawa E

机构信息

Department of Biochemistry, Medical College of Miyazaki, Japan.

出版信息

J Clin Lab Anal. 1992;6(2):105-12. doi: 10.1002/jcla.1860060206.

Abstract

A sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for (anti-human T-cell leukemia virus type I) IgG (anti-HTLV-I IgG) in serum using a synthetic peptide, Cys-env gp46(188-224) of HTLV-I, is described. Anti-HTLV-I IgG in test serum, which had been incubated with excess of inactive beta-D-galactosidase to eliminate interference by anti-beta-D-galactosidase antibodies, was reacted simultaneously with 2,4-dinitrophenyl-bovine serum albumin-Cys-env gp46 (188-224) conjugate and Cys-env gp46 (188-224)-beta-D-galactosidase conjugate. The complex formed consisting of the three components was trapped onto polystyrene balls coated with affinity-purified (anti-2,4-dinitrophenyl group) IgG. After washing to eliminate nonspecific IgG in the test serum and excess of the beta-D-galactosidase conjugate, the complex was eluted from the polystyrene balls with epsilon N-2,4-dinitrophenyl-L-lysine and transferred to polystyrene balls coated with affinity-purified (anti-human IgG gamma-chain) IgG. beta-D-Galactosidase activity bound to the (anti-human IgG gamma-chain) IgG-coated polystyrene balls was assayed by fluorometry. This assay was more sensitive and useful than the immune complex transfer enzyme immunoassay using Cys-Arg-env gp46(188-209) and other methods using HTLV-I as antigen.

摘要

描述了一种使用合成肽HTLV-I的Cys-env gp46(188 - 224)检测血清中抗人T细胞白血病病毒I型(anti-HTLV-I)IgG的灵敏酶免疫测定法(免疫复合物转移酶免疫测定法)。将检测血清中的抗HTLV-I IgG与过量的无活性β-D-半乳糖苷酶孵育以消除抗β-D-半乳糖苷酶抗体的干扰后,使其与2,4-二硝基苯基-牛血清白蛋白-Cys-env gp46(188 - 224)偶联物和Cys-env gp46(188 - 224)-β-D-半乳糖苷酶偶联物同时反应。由这三种成分形成的复合物被捕获到包被有亲和纯化的(抗2,4-二硝基苯基基团)IgG的聚苯乙烯球上。洗涤以消除检测血清中的非特异性IgG和过量的β-D-半乳糖苷酶偶联物后,用ε-N-2,4-二硝基苯基-L-赖氨酸从聚苯乙烯球上洗脱复合物,并转移到包被有亲和纯化的(抗人IgGγ链)IgG的聚苯乙烯球上。通过荧光法测定与(抗人IgGγ链)IgG包被的聚苯乙烯球结合的β-D-半乳糖苷酶活性。该测定法比使用Cys-Arg-env gp46(188 - 209)的免疫复合物转移酶免疫测定法和其他以HTLV-I为抗原的方法更灵敏且更有用。

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