Ligand Fishing: An Approach for the Discovery of Inhibitors from Complex Biological Mixtures.
TARGETING ENZYMES FOR PHARMACEUTICAL DEVELOPMENT: METHODS AND PROTOCOLS(2020)
摘要
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).
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关键词
Bioanalytical screening technique,GST enzyme natural inhibitors,Metal affinity chromatography,Screening active compounds
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