title: Development of Dissociation-Enhanced Lanthanide Fluoroimmunoassay for Measuring Leptin.
abstract: Development of a dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA) for measuring leptin, a satiety hormone of appetite control, was conducted in sandwich assay format exploiting a microplate immobilized with an anti-leptin antibody and another antibody raised against leptin and tagged with an europium chelate. In the leptin DELFIA of this study, amounts of antibody coated to the microplate and of the bioconjugate for the second immune reaction were optimized as 0.5 μg and 200 ng per well, respectively. When plotted in double-logarithmic scale, a linear relationship of y (log10 response signal) = 0.6023× (log10 leptin concentration) + 3.4084 (r(2) = 0.9646) was obtained at the leptin concentrations of 0.01─50 ng/mL with the limit of detection of 0.01 ng/mL. Individual leptin concentrations in various samples were well convergent to the calibration curve of the current assay. When applied to the measurement of leptin in a rat serum, the present assay was found quite effective and was competitive to a commercial sandwich-type ELISA.
authors.affiliation: Research Group of Bioprocess Engineering, Korea Food Research Institute, Seongnam, 13539, Republic of Korea. k9130sen@hanmail.net.
authors.forename: Namsoo
authors.initials: N
authors.lastname: Kim
authors.affiliation: Research Group of Bioprocess Engineering, Korea Food Research Institute, Seongnam, 13539, Republic of Korea.
authors.forename: So-Hee
authors.initials: SH
authors.lastname: Son
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005063:Europium / Q000737:chemistry*; D015200:Fluoroimmunoassay / Q000379:methods*; D028581:Lanthanoid Series Elements / Q000737:chemistry*; D020738:Leptin / Q000097:blood* / Q000276:immunology; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D000911:Antibodies, Monoclonal; D028581:Lanthanoid Series Elements; D020738:Leptin; D005063:Europium
keywords: Bioconjugate preparation; High-sensitivity detection; Immobilized-antibody microplate; Leptin; Reaction optimization