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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Hou X, Wang S, Zhang T, Ma J, Zhang J, Zhang Y, Lu W, He H, He L (2014) Recent advances in cell membrane chromatography for traditional Chinese medicines analysis. J Pharm Biomed Anal 101:141–150
Krogh A, Larsson BR, von Heijne G, Sonnhammer EL (2001) Predicting transmembrane protein topology with a hidden markov model: application to complete genomes. J Mol Biol 305:567–580
Overington JP, Al-Lazikani B, Hopkins AL (2006) How many drug targets are there. Nat Rev Drug Discov 5:993–996
He L, Wang S, Geng X (2001) Coating and fusing cell membranes onto a silica surface and their chromatographic characteristics. Chromatographia 54:71–76
He LC, Yang GD, Geng XD (1999) Enzymatic activity and chromatographic characteristics of the cell membrane immobilized on silica surface. Chin Sci Bull 44:826–831
He H, Han S, Zhang T, Zhang J, Wang S, Hou J (2012) Screening active compounds acting on the epidermal growth factor receptor from Radix scutellariae via cell membrane chromatography online coupled with HPLC/MS. J Pharm Biomed Anal 62:196–202
Wang L, Ren J, Sun M, Wang S (2010) A combined cell membrane chromatography and online HPLC/MS method for screening compounds from Radix Caulophylli acting on the human α1A-adrenoceptor. J Pharm Biomed Anal 51:1032–1036
Li M, Wang S, Zhang Y, He L (2010) An online coupled cell membrane chromatography with LC/MS method for screening compounds from Aconitum carmichaeli Debx. acting on VEGFR-2. J Pharm Biomed Anal 53:1063–1069
Tang C, Liu ZS, Qin N, Xu L, Duan HQ (2013) Novel cell membrane capillary chromatography for screening active compounds from natural products. Chromatograpia 76:697–701
Ding X, Chen X, Cao Y, Jia D, Wang D, Zhu Z, Zhang J, Hong Z, Chai Y (2014) Quality improvements of cell membrane chromatographic column. J Chromatogr A 1359:330–335
Tang C, Wu XD, Yu YM, Duan H, Zhou J, Xu L (2016) Cell extraction combined with off-line HPLC for screening active compounds from Coptis chinensis. Biomed Chromatogr 30:658–662
Dong ZB, Li SP, Hong M, Zhu Q (2005) Hypothesis of potential active components in Angelica sinensis by using biomembrane extraction and high performance liquid chromatography. J Pharm Biomed Anal 38:664–669
Zhuo R, Liu H, Liu N, Wang Y (2016) Ligand fishing: a remarkable strategy for discovering bioactive compounds from complex mixture of natural products. Molecules 21:1516–1531
de Moraes MC, Santos JB, Dos Anjos DM, Rangel LP, Vieira TC, Moaddel R, da Silva JL (2015) Prion protein-coated magnetic beads: synthesis, characterization and development of a new ligands screening method. J Chromatogr A 1379:1–8
Moaddel R, Marszałł MP, Bighi F, Yang Q, Duan X, Wainer IW (2007) Automated ligand fishing using human serum albumin-coated magnetic beads. Anal Chem 79:5414–5417
McFadden MJ, Junop MS, Brennan JD (2010) Magnetic “fishing” assay to screen small-molecule mixtures for modulators of protein-protein interactions. Anal Chem 82:9850–9857
Pochet L, Heus F, Jonker N, Lingeman H, Smit AB (2011) Online magnetic bead based dynamic protein affinity selection coupled to LC–MS for the screening of acetylcholine binding protein ligands. J Chromatogr B 879:1781–1788
Wubshet SG, Brighente IM, Moaddel R, Staerk D (2015) Magnetic ligand fishing as a targeting tool for HPLC–HRMS–SPE–NMR: α-glucosidase inhibitory ligands and alkylresorcinol glycosides from Eugenia catharinae. J Nat Prod 78:2657–2665
Zhang F, Zhao X, Xu B, Cheng S, Tang C, Duan H, Xiao X, Du W, Xu L (2016) 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
Marszałł MP, Moaddel R, Kole S, Gandhari M, Bernier M, Wainer IW (2008) Ligand and protein fishing with heat shock protein coated magnetic beads. Anal Chem 80:7571–7575
Qing LS, Xue Y, Deng WL, Liao X, Xu XM, Li BG, Liu YM (2011) Ligand fishing with functionalized magnetic nanoparticles coupled with mass spectrometry for herbal medicine analysis. Anal Bioanal Chem 399:1223–1231
Du H, He J, Wang S, He L (2010) Investigation of calcium antagonist–l-type calcium channel interactions by a vascular smooth muscle cell membrane chromatography method. Anal Bioanal Chem 397:1947–1953
Xie Y, Chen Y, Lin M, Wen J, Fan G, Wu Y (2007) High-performance liquid chromatographic method for the determination and pharmacokinetic study of oxypeucedanin hydrate and byak-angelicin after oral administration of Angelica dahurica extracts in mongrel dog plasma. J Pharm Biomed Anal 44:166–172
He JY, Zhang W, He LC, Cao YX (2007) Imperatorin induces vasodilatation possibly via inhibiting voltage dependent calcium channel and receptor-mediated Ca2+ influx and release. Eur J Pharmacol 573:170–175
Li B, Zhang X, Wang J, Zhang L, Gao BW, Shi SP, Wang XH, Li J, Tu PF (2014) Simultaneous characterisation of fifty coumarins from the roots of Angelica dahurica by off-line two-dimensional high-performance liquid chromatography coupled with electrospray ionisation tandem mass spectrometry. Phytochem Anal 25:229–240
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.
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Ethical approval
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.
Rights and permissions
About this article
Cite this article
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
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10337-017-3370-7