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Ligand Fishing with Cellular Membrane-Coated Magnetic Beads: A New Method for the Screening of Potentially Active Compounds from Natural Products

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Abstract

Ligand fishing with target biomolecule-immobilized magnetic beads (MBs) has been established and developed for nearly 10 years. Advantages of this technique, such as the ease of operation, associated with a diversity of automated online approaches, make it a valuable tool for affinity studies. However, transmembrane proteins have not been used as the target biomolecules in the assay, since they are usually not available in a purified and bioactive form. In addition, few publications have reported the use of this method for screening active compounds derived from natural products. In this work, for the first time, cellular membrane-coated MBs, which to a large extent maintain the activity of the transmembrane proteins, were used for the fishing assay. We demonstrated application of red blood cell membrane-coated MBs for fishing potential active components from a natural product (Angelica dahurica). The potential active compounds, such as imperatorin, bergapten, and pabulenol, were detected. The result correlated well with cell membrane chromatography (CMC) coupled with HPLC. Comparisons of the developed MBs fishing assay with the CMC method showed the noteworthy advantages of the fishing technique regarding the consumption of cellular membranes, buffers as well as length of operation time.

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Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (Nos. 81303191 and 81402889), Postdoctoral Foundation of China (No. 2012M510759), and the Natural Science Foundation of Tianjin (No. 14JCYBJC24300). We would like to thank Prof. L. He and Prof. S. Wang at Xi’an Jiaotong University for their guidance on CMC experiments carried out in this work. We would also like to thank Prof. Hongquan Duan of Tianjin Medical University for his support on this work.

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Authors and Affiliations

  1. Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics (Theranostics), School of Pharmacy, Tianjin Medical University, Tianjin, 300070, China

    Cheng Tang, Ruizhi Mao, Yaming Yu, Liang Xu & Yanwen Zhang

  2. Research Center of Basic Medical Sciences, Tianjin Medical University, Tianjin, 300070, China

    Cheng Tang, Ruizhi Mao, Yaming Yu & Liang Xu

  3. Tianjin Medical College, Tianjin, 300222, China

    Fang Liu, Liang Xu & Yanwen Zhang

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  1. Cheng Tang
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  2. Ruizhi Mao
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  3. Fang Liu
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  4. Yaming Yu
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  5. Liang Xu
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  6. Yanwen Zhang
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Correspondence to Liang Xu or Yanwen Zhang.

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All animal experiments have been approved by the Administrative Committee of Experimental Animal Care and Use of Tianjin Medical University and, furthermore, conformed to the guidelines set by the National Institute of Health on the ethical use of animals.

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Tang, C., Mao, R., Liu, F. et al. Ligand Fishing with Cellular Membrane-Coated Magnetic Beads: A New Method for the Screening of Potentially Active Compounds from Natural Products. Chromatographia 80, 1517–1525 (2017). https://doi.org/10.1007/s10337-017-3370-7

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  • DOI: https://doi.org/10.1007/s10337-017-3370-7

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