Subattomole detection of adiponectin in urine by ultrasensitive ELISA coupled with thio-NAD cycling

Mika Morikawa, Rina Naito, Koichi Mita, Satoshi Watabe, Kazunari Nakaishi, Teruki Yoshimura, Toshiaki Miura, Seiichi Hashida, Etsuro Ito*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)

Abstract

Adiponectin is a hormone secreted from adipocytes, and it demonstrates antidiabetic, anti-atherosclerotic, antiobesity and anti-inflammatory effects. However, the patterns of change in urinary adiponectin levels in various diseases remain unknown, because only trace amounts of the hormone are present in urine. In the present study, we applied an ultrasensitive ELISA coupled with thio-NAD cycling to measure urinary adiponectin levels. Spike-and-recovery tests using urine confirmed the reliability of our ultrasensitive ELISA. The limit of detection for adiponectin in urine was 2.3×10–19 moles/assay (1.4 pg/mL). The urinary adiponectin concentration ranged between 0.04 and 5.82 ng/mL in healthy subjects. The pilot study showed that the urinary adiponectin levels, which were corrected by the creatinine concentration, were 0.73±0.50 (ng/mg creatinine, N=6) for healthy subjects, versus 12.02±3.85 (ng/mg creatinine, N=3) for patients with diabetes mellitus (DM). That is, the urinary adiponectin levels were higher (P<0.05) in DM patients than in healthy subjects. Further, these urinary adiponectin levels tended to increase with the progression of DM accompanied with nephropathy. Our method is thus expected to provide a simple, rapid and reasonably priced test for noninvasive monitoring of the progression of DM without the requirement of special tools.

Original languageEnglish
Pages (from-to)79-86
Number of pages8
JournalBiophysics and physicobiology
Volume12
DOIs
Publication statusPublished - 2015
Externally publishedYes

Keywords

  • adiponectin
  • diabetes
  • ELISA
  • nephropathy
  • thio-NAD cycling
  • urine

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Physiology
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

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