One-step enzyme-linked immunosorbent assay (ELISA) for measurement of serum free leptin.
In man, circulating leptin levels are increased with obesity and are regulated by a complex of hormonal, feeding and body-weight changes. Accurate and precise methods to quantitate circulating serum free leptin (f-leptin) concentrations are needed for physiological and clinical studies. We developed a one-step enzyme immunoassay to measure human f-leptin in serum. The detection limit was 0.40 ng/ml. The recovery of leptin added to serum was 90.8-102.8%. The within-run and between-day coefficients of variation (C.V.) ranged from 2.8 to 7.7 and 5.7 to 9.7%, respectively, and the immunoassay had an overall recovery rate for serial dilution in the range of 94. 0-109.9%. Measured serum f-leptin concentrations in 201 adults correlated (r=0.449, P<0.001) directly with body mass index (BMI kg/m(2)), particularly when results were separated by gender (r=0. 709 for male, P<0.001; r=0.643 for female, P<0.001). We conclude that this one-step enzyme immunoassay is accurate for measuring f-leptin in human serum.
Kimura
E
E
Research Division, Japan Clinical Laboratories, Inc., 16-10, Ohashibe, Kumiyama, Kuze, Kyoto, Japan.
Matsumoto
K
K
Samori
T
T
Kato
S
S
Kawahara
T
T
D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000918:Antibody Specificity; D015992:Body Mass Index; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D020738:Leptin / Q000032:analysis*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009765:Obesity / Q000097:blood; D011786:Quality Control; D012016:Reference Values; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D020738:Leptin