title: Preparation and characterization of micro-cell membrane chromatographic column with silica-based porous layer open tubular capillary as cellular membrane carrier.
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. Graphical Abstract Comparision of mCMC chromatograms and SEM images between bare capillary and PLOT capillary.
authors.affiliation: Tianjin Huanhu Hospital, Tianjin, 300060, China.|Tianjin Key Laboratory of Cerebral Vascular and Neurodegenerative Diseases, Tianjin, 300060, China.
authors.forename: Fugeng
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics (Theranostics), School of Pharmacy, Tianjin Medical University, Tianjin, 300070, China.
authors.forename: Xinchao
authors.initials: X
authors.lastname: Zhao
authors.affiliation: School of Public Health, Tianjin Medical University, Tianjin, 300070, China.
authors.forename: Bei
authors.initials: B
authors.lastname: Xu
authors.affiliation: Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics (Theranostics), School of Pharmacy, Tianjin Medical University, Tianjin, 300070, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Cheng
authors.affiliation: Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics (Theranostics), School of Pharmacy, Tianjin Medical University, Tianjin, 300070, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Tang
authors.affiliation: Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics (Theranostics), School of Pharmacy, Tianjin Medical University, Tianjin, 300070, China.|Research Center of Basic Medical Sciences, Tianjin Medical University, Tianjin, 300070, China.
authors.forename: Hongquan
authors.initials: H
authors.lastname: Duan
authors.affiliation: Tianjin University of Traditional Chinese Medicine, Tianjin, 300193, China.
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Xiao
authors.affiliation: Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin, 300150, China.
authors.forename: Wuxun
authors.initials: W
authors.lastname: Du
authors.affiliation: Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics (Theranostics), School of Pharmacy, Tianjin Medical University, Tianjin, 300070, China. xuliang@tmu.edu.cn.
authors.forename: Liang
authors.initials: L
authors.lastname: Xu
mesh_terms: D000818:Animals; D002462:Cell Membrane; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008565:Membrane Proteins / Q000032:analysis*; D012822:Silicon Dioxide / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
chemical_list: D008565:Membrane Proteins; D012822:Silicon Dioxide
keywords: Affinity; Cell membrane chromatography; Micro-cell membrane chromatography; Porous layer open tubular capillary; Red blood cell