Kugimiya Akimitsu, Fukada Rie
Center for Industry and Public Relations, Hiroshima City University, 3-8-24 Senda-machi, Naka-ku, Hiroshima, 730-0052, Japan,
Appl Biochem Biotechnol. 2015 Jun;176(4):1195-202. doi: 10.1007/s12010-015-1639-6. Epub 2015 May 3.
Analysis of the concentration of free amino acids in biological samples is useful in clinical diagnostics. However, currently available methods are time consuming, potentially delaying diagnosis. Therefore, the development of more rapid analytical tools is needed. In this study, a chemiluminescence detection method for amino acids was developed, and the conditions for the enzyme reaction and assay were examined. For the recognition of each amino acid (here, serine, proline, glycine, asparagine, leucine, and histidine), the corresponding aminoacyl-tRNA synthetase (aaRS) was employed, and multiple enzymatic reactions were combined with a luminol chemiluminescence reaction. This method provided selective quantification from 1 to 20 μM for serine, proline, glycine, and leucine; 1 to 60 μM for asparagine; and 1 to 150 μM for histidine. This assay, which utilized aaRSs for the detection of amino acids, could be useful for simple and rapid analysis of amino acids in clinical diagnostics.
分析生物样品中游离氨基酸的浓度在临床诊断中很有用。然而,目前可用的方法耗时较长,可能会延迟诊断。因此,需要开发更快速的分析工具。在本研究中,开发了一种用于氨基酸的化学发光检测方法,并研究了酶反应和测定的条件。为了识别每种氨基酸(此处为丝氨酸、脯氨酸、甘氨酸、天冬酰胺、亮氨酸和组氨酸),使用了相应的氨酰 - tRNA合成酶(aaRS),并将多个酶反应与鲁米诺化学发光反应相结合。该方法对丝氨酸、脯氨酸、甘氨酸和亮氨酸的选择性定量范围为1至20μM;对天冬酰胺为1至60μM;对组氨酸为1至150μM。这种利用aaRS检测氨基酸的测定方法可能有助于临床诊断中氨基酸的简单快速分析。