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.
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.
Kohno
T
T
Department of Biochemistry, Medical College of Miyazaki, Japan.
Sakoda
I
I
Ishikawa
E
E
D000595:Amino Acid Sequence; D005069:Evaluation Studies as Topic; D015481:HTLV-I Antibodies / Q000097:blood*; D015486:HTLV-I Antigens / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D007124:Immunoenzyme Techniques / Q000706:statistics & numerical data; D007074:Immunoglobulin G / Q000097:blood*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology*; D012680:Sensitivity and Specificity
C076957:Cys-env gp46(188-224); D015481:HTLV-I Antibodies; D015486:HTLV-I Antigens; D007074:Immunoglobulin G; D010446:Peptide Fragments