Development of a sensitive ELISA for human leptin, using monoclonal antibodies.
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.
R & D and Manufacturing Department for Diagnostics, Shionogi Co., Ltd., Osaka, Japan.
K
K
Imagawa
Y
Y
Matsumoto
Y
Y
Numata
A
A
Morita
S
S
Kikuoka
M
M
Tamaki
C
C
Higashikubo
T
T
Tsuji
K
K
Sasakura
H
H
Teraoka
H
H
Masuzaki
K
K
Hosoda
Y
Y
Ogawa
K
K
Nakao
D000273:Adipose Tissue / Q000378:metabolism*; D000818:Animals; D000856:Anorexia Nervosa / Q000097:blood; D000911:Antibodies, Monoclonal / Q000276:immunology; D002555:Cerebrospinal Fluid / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D020738:Leptin; D051379:Mice; D008810:Mice, Inbred C57BL; D011506:Proteins / Q000032:analysis* / Q000276:immunology; D011863:Radioimmunoassay; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D020738:Leptin; D011506:Proteins; D011994:Recombinant Proteins