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Microfluidic analysis of serine levels using seryl-tRNA synthetase coupled with spectrophotometric detection
Akimitsu Kugimiya1, Emi Matsuzaki
1Center for Industry and Public Relations, Hiroshima City University, 3-8-24 Senda-machi, Naka-ku, Hiroshima, 730-0052, Japan, kugimiya@hiroshima-cu.ac.jp.
Abstract:
The measurement of amino acid content is useful for the diagnosis of several types of diseases, including cancer and diabetes. In this study, a microfluidic method for the analysis of serine using enzymatic reactions coupled with spectrophotometric detection was developed. The assay system has some advantages in the analytical field, such as the ability to detect small amounts of analyte and reaction solution and a rapid and efficient reaction. For the specific detection of serine, seryl-tRNA synthetase was coupled with the generation of hydrogen peroxide, which was then detected by the Trinder reagent spectrophotometric method. Seryl- and other aminoacyl-tRNA synthetases are involved in the biosynthesis of peptides and proteins in the human body and should allow precise recognition of the corresponding amino acids. This approach provided selective quantitation of up to 250 μM serine in 100 mM Tris-HCl buffer (pH 8.0) in a semiautomatic system.
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