title: Chemiluminescent enzyme immunoassay for measuring leptin.
abstract: Leptin is one of the representative adipocyte-derived protein hormones. Measuring the serum leptin concentration gives an important index for preventing and treating diabetes mellitus and other diseases. We constructed in this study a chemiluminescent enzyme immunoassay (CLEIA) for measuring leptin by using the anti-leptin polyclonal antibody and alkaline phosphatase (ALP). The method applies the IgG-conjugated ferrite particle to capture leptin in a sample and the ALP-conjugated Fab fragment to detect the captured leptin. We tested Block ace, CE510, and bovine serum albumin (BSA) for their abilities to block non-specific binding of ALP-conjugated anti-leptin Fab to the ferrite particle and found BSA to be the most effective. The measurable range with this ELISA for leptin was 0.1-1.0 pg/mL of leptin and the detection limit (blank+2SD) was 0.1 pg/mL of leptin. These results demonstrate sufficient sensitivity with our system to measure the serum leptin concentration and its clinical usefulness. The results also suggest that a sensitive enzyme immunoassay can be constructed by using only one polyclonal antibody.
authors.affiliation: Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Sekiguchi
authors.forename: Hideki
authors.initials: H
authors.lastname: Kohno
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Yasukawa
authors.forename: Kuniyo
authors.initials: K
authors.lastname: Inouye
mesh_terms: D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods*; D002138:Calibration; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D007202:Indicators and Reagents / Q000737:chemistry; D020738:Leptin / Q000097:blood* / Q000276:immunology; D008163:Luminescent Measurements / Q000379:methods*; D008297:Male
chemical_list: D007202:Indicators and Reagents; D020738:Leptin