An enzyme immunoassay for human lymphotoxin.
A highly sensitive and specific enzyme immunoassay (EIA) for human lymphotoxin (hLT) has been developed. The assay is based upon a sandwich system employing two kinds of anti-hLT antibodies with neutralizing activity. One of them was mouse monoclonal antibody raised against Escherichia coli-derived recombinant hLT with a deletion of 20 amino-terminal amino acids and used as labelled antibody. The other was rabbit antibody raised against the carboxyl-terminal portion of hLT and used as solid-phase antibody. The EIA employing such a combination was able to detect less than 50 pg/ml of hLT, showing that this method was approximately 5-10 times higher sensitivity than the conventional bioassay employing L929 cell-lysis. The mean recovery of hLT added to serum specimens was 101% and the coefficients of variation were 3.3-7.8% (intra-assay) and 2.9-17.2% (interassay). There was a good correlation between the present EIA and the bioassay (r = 0.93).
Tada
H
H
Central Research Division, Takeda Chemical Industries, Ltd., Osaka, Japan.
Toyoda
Y
Y
Okazaki
K
K
Nakata
M
M
Iwasa
S
S
D000818:Animals; D000911:Antibodies, Monoclonal; D001681:Biological Assay; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007124:Immunoenzyme Techniques; D008233:Lymphotoxin-alpha / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D008233:Lymphotoxin-alpha; D011994:Recombinant Proteins