Novel and sensitive enzyme immunoassay (immune-complex-transfer enzyme immunoassay) for antihuman T cell leukemia virus type 1 IgG in human serum using recombinant gag-env hybrid protein as antigen.
A novel and sensitive enzyme immunoassay (immune-complex-transfer enzyme immunoassay) for antihuman T cell leukemia virus type 1 IgG (anti-HTLV-1 IgG) in human serum using recombinant gag(14-139)-env-(197-295) hybrid protein is described. Anti-HTLV-1 IgG in test serum was reacted with dinitrophenyl biotinyl bovine serum albumin-recombinant gag-env hybrid protein conjugate. The complex formed was trapped onto polystyrene balls coated with affinity-purified antidinitrophenyl group IgG. After washing to eliminate nonspecific IgG in the test serum, the complex was eluted from the polystyrene balls with dinitrophenyl-L-lysine and transferred to polystyrene balls coated with streptavidin. After washing, anti-HTLV-1 IgG in the complex trapped onto the streptavidin-coated polystyrene balls was reacted with antihuman IgG gamma-chain Fab'-peroxidase conjugate. Peroxidase activity bound to the streptavidin-coated polystyrene balls was assayed by fluorometry. By transfer of the complex, the nonspecific binding of nonspecific human IgG was considerably reduced, and the detection limit of anti-HTLV-1 IgG in serum was lowered 30-300-fold compared with that by Western blotting, gelatin particle agglutination, and the conventional enzyme immunoassay, in which a recombinant gag-env hybrid protein-coated polystyrene ball was incubated with test serum and, after washing, with antihuman IgG gamma-chain Fab'-peroxidase conjugate. Usefulness of the immune-complex-transfer enzyme immunoassay was demonstrated using 271 serum samples.
Kohno
H
H
Fuji Research Laboratories, Kyowa Medex Co., Ltd., Shizuoka, Japan.
Kohno
T
T
Sakoda
I
I
Ishikawa
E
E
D015153:Blotting, Western; D004140:Dinitrophenols; D015486:HTLV-I Antigens / Q000097:blood*; D015368:Human T-lymphotropic virus 1 / Q000032:analysis*; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000032:analysis*; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D012710:Serum Albumin, Bovine; D014764:Viral Proteins / Q000276:immunology*
D004140:Dinitrophenols; D015486:HTLV-I Antigens; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C034638:dinitrophenyl-bovine serum albumin; D012710:Serum Albumin, Bovine