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Ligand Fishing: An Approach for the Discovery of Inhibitors from Complex Biological Mixtures

  • Evangelia G. Chronopoulou
  • Christina Varotsou
  • Nikolaos Georgakis
  • Georgios Premetis
  • Elisavet Ioannou
  • Nikolaos E. LabrouEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2089)

Abstract

Ligand fishing is a convenient bioanalytical screening method that is based on the affinity selection of a ligand from a complex biological sample by an immobilized receptor. It is a versatile affinity-based screening approach and it has found application in multiple interacting pairs such as enzyme-inhibitor/activator, antigen-antibody, receptor-ligand, and protein-protein. Important parameters that affect the successful operation of the method are the high specificity and strong binding affinity of the interacting pair (e.g., enzyme-ligand complex) and the elution of the bound ligand from the complex. This chapter provides protocols for the synthesis of affinity adsorbent and its application in off-line ligand-fishing procedure for a 6His-tagged glutathione transferase (GST).

Key words

Metal affinity chromatography Screening active compounds Bioanalytical screening technique GST enzyme natural inhibitors 

Notes

Acknowledgments

E.G.C. thanks IKY Scholarship Programs that is co-financed by Greece and the European Union (European Social Fund- ESF) through the Operational Programme «Human Resources Development, Education and Lifelong Learning» in the context of the project “Reinforcement of Postdoctoral Researchers” (MIS-5001552), implemented by the State Scholarships Foundation (IKY).

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Copyright information

© Springer Science+Business Media, LLC 2020

Authors and Affiliations

  • Evangelia G. Chronopoulou
    • 1
  • Christina Varotsou
    • 1
  • Nikolaos Georgakis
    • 1
  • Georgios Premetis
    • 1
  • Elisavet Ioannou
    • 1
  • Nikolaos E. Labrou
    • 1
    Email author
  1. 1.Laboratory of Enzyme Technology, Department of BiotechnologySchool of Applied Biology and Biotechnology, Agricultural University of AthensAthensGreece

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