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Preparation and characterization of micro-cell membrane chromatographic column with silica-based porous layer open tubular capillary as cellular membrane carrier

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Abstract

Cell membrane chromatography (CMC) is a powerful tool to study membrane protein interactions and to screen active compounds extracted from natural products. Unfortunately, a large amount of cells are typically required for column preparation in order to carry out analyses in an efficient manner. Micro-CMC (mCMC) has recently been developed by using a silica capillary as a membrane carrier. However, a reduced retention of analytes is generally associated with mCMC mostly due to a low ligand (cellular membrane) capacity. To solve this common problem, in this work a silica-based porous layer open tubular (PLOT) capillary was fabricated and, to the best of our knowledge, for the first time applied to mCMC. The mCMC column was prepared by physical adsorption of rabbit red blood cell (rRBC) membranes onto the inner surface of the PLOT capillary. The effects of the PLOT capillaries fabricated by different feed compositions, on the immobilization amount of cellular membranes (represented by the fluorescence intensity of the capillary immobilized with fluorescein isothiocyanate isomer-labeled cellular membranes) and on the dynamic binding capacity (DBC) of verapamil (VP, a widely used calcium antagonist which specific interacts with L-type calcium channel proteins located on cellular membrane of rRBC) have been systematically investigated. The fluorescence intensity of the mCMC column when combined with the PLOT capillary was found to be more than five times higher than the intensity using a bare capillary. This intriguing result indicates that the PLOT capillary exhibits a higher cellular membrane capacity. The DBC of VP in the PLOT column was found to be more than nine times higher than that in the bare capillary. An rRBC/CMC column was also prepared for comparative studies. As a result, mCMC provides similar chromatographic retention factors and stability with common CMC; however, the cellular membrane consumption for mCMC was found to be more than 460 times lower than that for CMC.

Comparision of mCMC chromatograms and SEM images between bare capillary and PLOT capillary

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Acknowledgment

This work was financially supported by the National Natural Science Foundation of China (No. 81402889 and No. 81303191) and the Natural Science Foundation of Tianjin (No. 14JCYBJC24300). We would also like to thank Prof. L. He and Prof. S. Wang at Xi’an Jiaotong University for their valuable guidance on this work.

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Authors and Affiliations

  1. Tianjin Huanhu Hospital, Tianjin, 300060, China

    Fugeng Zhang

  2. Tianjin Key Laboratory of Cerebral Vascular and Neurodegenerative Diseases, Tianjin, 300060, China

    Fugeng Zhang

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

    Xinchao Zhao, Shuai Cheng, Cheng Tang, Hongquan Duan & Liang Xu

  4. School of Public Health, Tianjin Medical University, Tianjin, 300070, China

    Bei Xu

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

    Hongquan Duan

  6. Tianjin University of Traditional Chinese Medicine, Tianjin, 300193, China

    Xuefeng Xiao

  7. Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin, 300150, China

    Wuxun Du

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  1. Fugeng Zhang
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  2. Xinchao Zhao
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  9. Liang Xu
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Correspondence to Liang Xu.

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The 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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Zhang, F., Zhao, X., Xu, B. et al. Preparation and characterization of micro-cell membrane chromatographic column with silica-based porous layer open tubular capillary as cellular membrane carrier. Anal Bioanal Chem 408, 2441–2448 (2016). https://doi.org/10.1007/s00216-016-9339-9

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  • DOI: https://doi.org/10.1007/s00216-016-9339-9

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