High-throughput determination of free d-aspartic acid in mammals by enzyme immunoassay using specific monoclonal antibody

Tomohiro Ohgusu, Kenji Hamase, Hiroyuki Tanaka, Yukihiro Shoyama, Kiyoshi Zaitsu

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

A method for rapid determination of free d-aspartic acid (d-Asp) in mammals has been established using a highly specific mouse monoclonal antibody against d-Asp for the first time. An anti-d-Asp monoclonal antibody was obtained by the immunization of bovine-serum-albumin-conjugated d-Asp to BALB/c mice. The obtained antibody has a high specificity toward d-Asp but shows a slight cross-reactivity to all other d- and l- amino acids including l-Asp. The calibration range of the competitive enzyme linked immunosorbent assay (ELISA) is 0.016-16 μmol/mL d-Asp in rat serum samples. The precisions of this method were evaluated by inter-plate and intraplate assays, and the relative standard deviation values were 4.8% and 4.5%, respectively. The values of d-Asp determined by the present ELISA have a good correlation to those determined by high-performance liquid chromatography with the correlation coefficient of 0.963. Using this ELISA, the time course of d-Asp in the rat serum after intravenous administration was successfully demonstrated. The present method provides a simple and high-throughput determination of d-Asp in mammals, and is a useful tool for clarifying the physiological roles and diagnostic values of this d-amino acid.

Original languageEnglish
Pages (from-to)15-20
Number of pages6
JournalAnalytical Biochemistry
Volume357
Issue number1
DOIs
Publication statusPublished - Oct 1 2006

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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