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Clinical Chemistry 44: 2165-2171, 1998;
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(Clinical Chemistry. 1998;44:2165-2171.)
© 1998 American Association for Clinical Chemistry, Inc.

Endocrinology and Metabolism

Development of a sensitive ELISA for human leptin, using monoclonal antibodies

Keiichi Imagawa1, Yayoi Matsumoto1, Yoshito Numata1,a, Atsushi Morita1, Shino Kikuoka1, Mikio Tamaki1, Chie Higashikubo1, Tetsuo Tsuji1, Kazuyuki Sasakura1, Hiroshi Teraoka1, Hiroaki Masuzaki2, Kiminori Hosoda2, Yoshihiro Ogawa2, and Kazuwa Nakao2

1 R & D and Manufacturing Department for Diagnostics, Diagnostic Science Division, Shionogi Co., Ltd., 2-5-1 Mishima, Settsu-shi, Osaka 566-0022, Japan.

2 Department of Medicine and Clinical Science, Kyoto University Graduate School of Medicine, 54 Shogoin Kawahara-cho, Sakyo-ku, Kyoto 606-8507, Japan.
a Author for correspondence. Fax 81-6-319-4109; e-mail yoshito.numata{at}shionogi.co.jp.

A new, sensitive ELISA for human leptin in plasma and cerebrospinal fluid (CSF) was developed, using monoclonal antibodies. The lower limit of detection of this ELISA was 0.78 pg/assay. Both intra- and interassay imprecision values were <7%. The dilution curves of plasma and CSF showed good linearity, and the recovery was 83.2–95.6%. There was good correlation between plasma leptin concentrations by the ELISA and a commercially available RIA (r = 0.99). Our ELISA is advantageous because it does not require radioisotopes, it produces results in hours rather than days, and more importantly, it improves on the detection limit and plasma interference of the RIA kit. The new ELISA enables measurement of low concentrations of leptin, as are seen in CSF and in plasma of patients with anorexia nervosa.






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Copyright © 1998 by the American Association for Clinical Chemistry.