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Sandwich enzyme immunoassay for rat transferrin with two monoclonal antibodies and its application
T Matsumoto1, H Shima, T Kishi
1Department of Biochemistry, Research Laboratory of Applied Biochemistry, Tanabe Seiyaku Company Ltd., Osaka, Japan.
The development of a sandwich enzyme immunoassay for rat transferrin with two monoclonal antibodies is described. Microtiter plates coated with one monoclonal antibody (15C2H3) were used, and captured transferrin was estimated with a horseradish peroxidase-conjugated Fab' fragment of another monoclonal antibody (22A06D2). In this assay, the measurable range is 5-150 ng/ml and the coefficients of variation within and between the assay series are 1.2-5.0 and 3.3-6.0%, respectively. Recovery was 101 +/- 9.7% when purified rat transferrin was added to rat plasma. No cross-reactivity with bovine, human, or mouse transferrin was shown. This assay for rat transferrin is a highly specific, sensitive, and expeditious method which may allow routine analysis of rat transferrin in blood or culture supernatants of rat hepatocytes.
The development of a sandwich enzyme immunoassay for rat transferrin with two monoclonal antibodies is described. Microtiter plates coated with one monoclonal antibody (15C2H3) were used, and captured transferrin was estimated with a horseradish peroxidase-conjugated Fab' fragment of another monoclonal antibody (22A06D2). In this assay, the measurable range is 5-150 ng/ml and the coefficients of variation within and between the assay series are 1.2-5.0 and 3.3-6.0%, respectively. Recovery was 101 +/- 9.7% when purified rat transferrin was added to rat plasma. No cross-reactivity with bovine, human, or mouse transferrin was shown. This assay for rat transferrin is a highly specific, sensitive, and expeditious method which may allow routine analysis of rat transferrin in blood or culture supernatants of rat hepatocytes.