Development of a Fluorescence-Linked Immunosorbent Assay for Baicalin

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• 作者：Wenchao Shan ; Jinjun Cheng ; Baoping Qu ; Jiayang Sai ; Hui Kong…
• 关键词：Baicalin ; Fluorescence ; linked immunosorbent assay ; Enzyme ; linked immunosorbent assay ; Traditional Chinese medicine ; Fluorescently labelled monoclonal antibody
• 刊名：Journal of Fluorescence
• 出版年：2015
• 出版时间：September 2015
• 年：2015
• 卷：25
• 期：5
• 页码：1371-1376
• 全文大小：346 KB
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• 作者单位：Wenchao Shan (1)
  Jinjun Cheng (1)
  Baoping Qu (2)
  Jiayang Sai (3)
  Hui Kong (2)
  Huihua Qu (4)
  Yan Zhao (2)
  Qingguo Wang (2)
  
  1. School of Chinese Materia Medica, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China
  2. School of Basic Medical Sciences, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China
  3. The third affiliated hospital, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China
  4. Center of Scientific Experiment, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, 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

Previously, we developed an indirect competitive enzyme-linked immunosorbent assay (icELISA) for baicalin (BAL) and used this assay to investigate the pharmacokinetics of BAL in mice. In this study, an anti-BAL monoclonal antibody (MAb) was purified by the caprylic acid method and then labelled with fluorescein isothiocyanate (FITC). Subsequently, an indirect competitive fluorescence-linked immunosorbent assay (icFLISA) was developed to detect baicalin (BAL) using FITC-labelled anti-BAL MAbs. Characterization of the assay demonstrated an effective BAL measurement range of 6.4 ng/mL to 500 μg/mL (R2--.997). The relative standard deviations (RSDs) for both intra-assay and inter-assay repeatability and precision were below 10 %. This icFLISA for BAL is simple, rapid and sensitive, with a 390-fold larger linear range and a 2-fold lower limit of detection (LOD) compared with the previously developed icELISA. We observed a strong correlation between the results determined by the icFLISA and icELISA methods. Overall, this study provides a useful method for detecting BAL in medicines, enabling in vivo visualization research. Keywords Baicalin Fluorescence-linked immunosorbent assay Enzyme-linked immunosorbent assay Traditional Chinese medicine Fluorescently labelled monoclonal antibody