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

被引:13
作者
Tang, Cheng [1 ,2 ]
Mao, Ruizhi [1 ,2 ]
Liu, Fang [3 ]
Yu, Yaming [1 ,2 ]
Xu, Liang [1 ,2 ,3 ]
Zhang, Yanwen [1 ,3 ]
机构
[1] Tianjin Med Univ, Sch Pharm, Dev Clin Therapeut & Diagnost Theranost, Tianjin Key Lab Technol Enabling, Tianjin 300070, Peoples R China
[2] Tianjin Med Univ, Res Ctr Basic Med Sci, Tianjin 300070, Peoples R China
[3] Tianjin Med Coll, Tianjin 300222, Peoples R China
基金
中国国家自然科学基金;
关键词
Fishing; Magnetic beads; Red blood cell; Membrane; Angelica dahurica; PERFORMANCE LIQUID-CHROMATOGRAPHY; ANGELICA-DAHURICA; SILICA SURFACE; EXTRACTION; CAPILLARY; RECEPTOR; COLUMN; HPLC;
D O I
10.1007/s10337-017-3370-7
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Ligand fishing with targetbiomolecule-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 membranecoated 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 membranecoated 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.
引用
收藏
页码:1517 / 1525
页数:9
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