RRC ID 42735
著者 Ohgusu T, Hamase K, Tanaka H, Shoyama Y, Zaitsu K.
タイトル High-throughput determination of free D-aspartic acid in mammals by enzyme immunoassay using specific monoclonal antibody.
ジャーナル Anal Biochem
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 micromol/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.
巻・号 357(1)
ページ 15-20
公開日 2006-10-1
DOI 10.1016/j.ab.2006.06.039
PII S0003-2697(06)00490-8
PMID 16920063
MeSH Amino Acid Sequence Animals Antibodies, Monoclonal / immunology* Aspartic Acid / blood* Aspartic Acid / chemistry Calibration Cattle Enzyme-Linked Immunosorbent Assay / methods* Female Injections, Intravenous Mice Mice, Inbred BALB C Molecular Sequence Data Ovalbumin / chemistry Rats Sensitivity and Specificity Serum Albumin, Bovine / chemistry Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Time Factors
IF 2.877
引用数 4
WOS 分野 CHEMISTRY, ANALYTICAL BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
ヒト・動物細胞 SP2/0-Ag14(RCB0209)