Development of Radioimmunoassay for Human Leptin

• Published in: Biochemical and biophysical research communications Vol. 221; no. 2; pp. 234 - 239
• Main Authors: Hosoda, Kiminori, Masuzaki, Hiroaki, Ogawa, Yoshihiro, Miyawaki, Takashi, Hiraoka, Junko, Hanaoka, Ikuko, Yasuno, Akiko, Nomura, Toshiyuki, Fujisawa, Yukio, Yoshimasa, Yasunao, Nishi, Shigeo, Yamori, Yukio, Nakao, Kazuwa
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 16.04.1996
• Subjects: Adipose Tissue - metabolism; Culture Media; DNA, Complementary; Humans; Leptin; obesity; Obesity - metabolism; proteins; Proteins - metabolism; Radioimmunoassay; radioimmunoassays; Recombinant Proteins - metabolism; structural genes
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1006/bbrc.1996.0579

Using recombinant human leptin, we have produced an antiserum for human leptin and developed a radioimmunoassay (RIA) specific and sensitive for human leptin. We detected leptin-like immunoreactivity (-LI) in culture media of adipose tissue from subcutaneous abdominal fat in human. The plasma human leptin-LI concentration in nonobese subjects (17.6 < body mass index (BMI) < 23.0) was 5.6 ± 1.3 (mean ± SE) ng/ml, while that in obese subjects (29.0 < BMI) was 43.0 ± 43.0 ± 9.4 (mean ± SE) ng/ml. In gel permeation chromatographic analyses, leptin-LI in culture media and plasma consisted of single component emerging at the elution position of recombinant human leptin. These findings indicate that leptin is secreted from the adipose tissue into the circulating blood as a large molecular form corresponding to recombinant leptin. The RIA developed in the present study will be a powerful tool to investigate the physiological and pathophysiological significance of leptin in human.