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 journalArticle

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

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Mammals
Immunoenzyme Techniques
Aspartic Acid
Immunosorbents
Assays
Enzyme-Linked Immunosorbent Assay
Monoclonal Antibodies
Throughput
Enzymes
Rats
Amino Acids
Bovine Serum Albumin
Immunization
Serum
Intravenous Administration
Calibration
High performance liquid chromatography
High Pressure Liquid Chromatography
Antibodies

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

High-throughput determination of free d-aspartic acid in mammals by enzyme immunoassay using specific monoclonal antibody. / Ohgusu, Tomohiro; Hamase, Kenji; Tanaka, Hiroyuki; Shoyama, Yukihiro; Zaitsu, Kiyoshi.

In: Analytical Biochemistry, Vol. 357, No. 1, 01.10.2006, p. 15-20.

Research output: Contribution to journalArticle

Ohgusu, Tomohiro ; Hamase, Kenji ; Tanaka, Hiroyuki ; Shoyama, Yukihiro ; Zaitsu, Kiyoshi. / High-throughput determination of free d-aspartic acid in mammals by enzyme immunoassay using specific monoclonal antibody. In: Analytical Biochemistry. 2006 ; Vol. 357, No. 1. pp. 15-20.
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