Detection of Dopamine and Serotonin by Competitive Enzyme-Linked Immunosorbent Assay

Su Min Namkung, Jeong Su Choi, Ji Hyang Park, Man Gil Yang, Min Woo Lee, Suhng Wook Kim
2017 Korean Journal of Clinical Laboratory Science  
1 고려대학교 대학원 의생명융합과학과, 2 서울대학교병원 의생명연구원 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 g/mL BSA conjugate on a 96-well surface, hormone and primary Ab, which are respectively
more » ... respectively diluted to different concentrations, were treated. Then, HRP-conjugated secondary Ab and TMB were added to measure absorbance. The regression equation and R 2 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, R 2 , 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, R 2 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. 서 론 도파민(dopamine, DA)과 세로토닌(serotonin, 5-Hydroxytryptamine, 5-HT)은 신경전달물질 및 호르몬으로서 체내에 미량 존재하지만 그 범위를 벗어날 경우 심각한 질병 상태를 초 래할 수 있다. 도파민은 모노아민에 속하는 카테콜아민 계열 신 경전달물질 중 하나로 전구체는 L-DOPA이고, 인간 뇌에서 노 르에피네프린으로 생합성된다. 혈액뇌장벽(Blood-Brain Barrier, BBB)을 통과하지 못 하기 때문에 뇌와 말초에서의 합성과 기능 은 독립적이다[1]. 혈관 확장 및 심박수와 혈압 증가, 배뇨량 증 가, 인슐린 생성 감소, 운동 조절, 동기부여, 보상기전 등의 기능 을 하며[2] 운동 조절 중에서도 자발적인 움직임에 중요한 기능 을 하기 때문에 부족할 경우 파킨슨병을 유발한다[3]. 도파민의 혈청 농도는 30 ng/L (0.195×10 −9 M) 미만이며, 24시간 뇨에 서의 농도는 26∼480 g/L (170∼3,319×10 −9 M)이다[4]. 혈 청 농도가 매우 낮기 때문에 검사 시 혈청보다는 24시간 뇨 검체
doi:10.15324/kjcls.2017.49.3.220 fatcat:ig53dz2225d4hmc22wg3qvy3ve