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Development of a Fluorescence-Linked Immunosorbent Assay for Baicalin

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Abstract

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 = 0.997). 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.

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Acknowledgments

This research was supported by the National Natural Science Foundation of China (81473338, 81373542), the National Key Basic Research Development Program (973 program, 2011CB505101) and the Classical Prescription Basic Research Team of Beijing University of Chinese Medicine. This manuscript has been thoroughly edited by a native English speaker from Elsevier Language Editing Services.

Conflict of Interest

The authors declare no conflicts of interest.

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    Authors and Affiliations

    1. School of Chinese Materia Medica, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China

      Wenchao Shan & Jinjun Cheng

    2. School of Basic Medical Sciences, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China

      Baoping Qu, Hui Kong, Yan Zhao & Qingguo Wang

    3. The third affiliated hospital, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China

      Jiayang Sai

    4. Center of Scientific Experiment, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China

      Huihua Qu

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    1. Wenchao Shan
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    2. Jinjun Cheng
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    3. Baoping Qu
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    4. Jiayang Sai
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    5. Hui Kong
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    7. Yan Zhao
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    8. Qingguo Wang
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    Corresponding author

    Correspondence to Huihua Qu.

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    Wenchao Shan and Jinjun Cheng contributed equally to this work.

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    Shan, W., Cheng, J., Qu, B. et al. Development of a Fluorescence-Linked Immunosorbent Assay for Baicalin. J Fluoresc 25, 1371–1376 (2015). https://doi.org/10.1007/s10895-015-1627-9

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    • DOI: https://doi.org/10.1007/s10895-015-1627-9

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