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

• Morikawa Mika

  R&D Headquarters, TAUNS Laboratories, Inc. Kagawa School of Pharmaceutical Sciences, Tokushima Bunri University

• Naito Rina

  Kagawa School of Pharmaceutical Sciences, Tokushima Bunri University

• Mita Koichi

  Faculty of Science and Engineering, Tokushima Bunri University

• Watabe Satoshi

  R&D Department, BL Co. Ltd.

• Nakaishi Kazunari

  R&D Headquarters, TAUNS Laboratories, Inc.

• Yoshimura Teruki

  Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido

• Miura Toshiaki

  Graduate School of Pharmaceutical Sciences, Hokkaido University

• Hashida Seiichi

  Institute for Health Sciences, Tokushima Bunri University

• Ito Etsuro

  Kagawa School of Pharmaceutical Sciences, Tokushima Bunri University

抄録

Adiponectin is a hormone secreted from adipocytes, and it demonstrates antidiabetic, anti-atherosclerotic, anti-obesity 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.

収録刊行物

• Biophysics and Physicobiology

  Biophysics and Physicobiology 12 (0), 79-86, 2015

  一般社団法人 日本生物物理学会

キーワード

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

詳細情報

• CRID: 1390001205764941824
• NII論文ID: 130005109683
• DOI: 10.2142/biophysico.12.0_79
• ISSN: 21894779
• Web Site: https://www.jstage.jst.go.jp/article/biophysico/12/0/12_79/_pdf
• 本文言語コード: en
• データソース種別:
  • JaLC
  • Crossref
  • CiNii Articles
  • KAKEN
• 抄録ライセンスフラグ: 使用不可