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KMID : 1101420170490030220
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Korean Journal of Clinical Laboratory Science
2017 Volume.49 No. 3 p.220 ~ p.226
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Detection of Dopamine and Serotonin by Competitive Enzyme-Linked Immunosorbent Assay
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Namkung Su-Min
Choi Jeong-Su
Park Ji-Hyang
Yang Man-Gil
Lee Min-Woo
Kim Suhng-Wook
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Abstract
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Dopamine (DA) and serotonin (5-Hydroxytryptamine, 5-HT) are neurotransmitters and hormones that exist in small amounts but have important role in the body. Serum and 24-hour urine are used as specimens, and are usually examined by HPLC-MS. In this study, we tried to detect DA and 5-HT by competitive ELISA using antigen-antibody (Ab) reaction. After immobilizing 5 μg/mL BSA conjugate on a 96-well surface, hormone and primary Ab, which are respectively diluted to different concentrations, were treated. Then, HRP-conjugated secondary Ab and TMB were added to measure absorbance. The regression equation and R2 value were calculated based on absorbance, and sensitivity of Ab to hormone as well as the correlation between hormone concentration and absorbance were determined. In DA ELISA, R2, the correlation between the concentration of hormone and absorbance, was the highest by 0.91 when anti-dopamine Ab was diluted 6,000 times and 7,000 times. In 5-HT ELISA, R2 was bigger than 0.90 in every concentration except 3,000 times and 6,000 times. Both DA and 5-HT were not effectively detected at low concentrations (less than 1.0×10?7 M); and because reference value of serum DA is lower than this, HPLC-MS was required to detect serum DA. However, competitive ELISA may be effective in detecting 24-hour urine DA, serum, and 24-hour 5-HT. Further studies are needed to detect hormones more accurately at lower concentrations.
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KEYWORD
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Competitive ELISA, Dopamine, Serotonin
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