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Original Articles

An Enzyme Immunoassay for Human Lymphotoxin

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Pages 93-105 | Published online: 23 Oct 2006
 

Abstract

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 aminoterminal 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).

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