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

被引:9
作者
KOHNO, T
SAKODA, I
ISHIKAWA, E
机构
[1] MIYAZAKI MED COLL,DEPT BIOCHEM,MIYAZAKI 88916,JAPAN
[2] JAPANESE RED CROSS,MIYAZAKI BLOOD CTR,MIYAZAKI,JAPAN
关键词
ANTIBODY; ADULT T-CELL LEUKEMIA; BETA-D-GALACTOSIDASE; WESTERN BLOTTING; GELATIN PARTICLE AGGLUTINATION; ELISA;
D O I
10.1002/jcla.1860060206
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
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.
引用
收藏
页码:105 / 112
页数:8
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