Development of Fluorescence-Linked Immunosorbent Assay for Paeoniflorin

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• 作者：Yan Zhao ; Huihua Qu ; Xueqian Wang ; Yue Zhang ; Jinjun Cheng…
• 关键词：Paeoniflorin (PF) ; Fluorescence ; Linked Immunosorbent Assay (FLISA) ; Enzyme ; Linked Immunosorbent Assay (ELISA) ; Determination ; Fluorescently labelled monoclonal antibody
• 刊名：Journal of Fluorescence
• 出版年：2015
• 出版时间：July 2015
• 年：2015
• 卷：25
• 期：4
• 页码：885-890
• 全文大小：329 KB
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• 作者单位：Yan Zhao (1)
  Huihua Qu (2)
  Xueqian Wang (1)
  Yue Zhang (1)
  Jinjun Cheng (3)
  Yan Zhao (1)
  Qingguo Wang (1)
  
  1. School of Basic Medical Sciences, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, China
  2. Scientific Research Experiment Center, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, China
  3. College of Traditional Chinese Pharmacy, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, China
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Biomedicine
  Biophysics and Biomedical Physics
  Biotechnology
  Biochemistry
  Analytical Chemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-4994

文摘

In this study, we developed a fluorescent immunoassay approach to detect paeoniflorin (PF) using a fluorescently labelled monoclonal antibody. The PF-specific antibody was purified by the caprylic acid-ammonium sulfate method and protein G Sepharose 4 Fast Flow column and?then labelled with fluorescein isothiocyanate (FITC). The FITC-labelled monoclonal antibody was highly specific for PF, with less than 0.076?% cross-reactivity to seven structurally related compounds. The FITC-labelled monoclonal antibody was then used to develop an indirect competitive enzyme-linked immunosorbent assay (icELISA) and indirect competitive fluorescence-linked immunosorbent assay (icFLISA), respectively. FLISA is simple, rapid and sensitive, with a 500-fold lower limit of detection (LOD) compared with conventional ELISA. Finally, using a variety of standards, FLISA was validated. We observed a strong correlation between the results determined by either FLISA or conventional HPLC for the quantification of PF levels (R2--.9927). Collectively, this study shows that the icFLISA method can be successfully applied for the detection and quantification of PF in medicines and biological samples.