Determination of half-maximal inhibitory concentration using biosensor-based protein interaction analysis

Anal Biochem. 2016 Sep 1:508:97-103. doi: 10.1016/j.ab.2016.06.025. Epub 2016 Jun 27.

Abstract

Half-maximal inhibitory concentration (IC50) is the most widely used and informative measure of a drug's efficacy. It indicates how much drug is needed to inhibit a biological process by half, thus providing a measure of potency of an antagonist drug in pharmacological research. Most approaches to determine IC50 of a pharmacological compound are based on assays that utilize whole cell systems. While they generally provide outstanding potency information, results can depend on the experimental cell line used and may not differentiate a compound's ability to inhibit specific interactions. Here we show using the secreted Transforming Growth Factor-β (TGF-β) family ligand BMP-4 and its receptors as example that surface plasmon resonance can be used to accurately determine IC50 values of individual ligand-receptor pairings. The molecular resolution achievable wih this approach can help distinguish inhibitors that specifically target individual complexes, or that can inhibit multiple functional interactions at the same time.

Keywords: BMP-4; Bone morphogenetic protein; Cerberus; IC(50); Inhibitor; SPR; Surface plasmon resonance; TGF-β.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Assay
  • Bone Morphogenetic Protein 4 / chemistry
  • Bone Morphogenetic Protein 4 / metabolism
  • Chemistry Techniques, Analytical / methods*
  • Humans
  • Inhibitory Concentration 50
  • Protein Binding
  • Receptors, Transforming Growth Factor beta / chemistry*
  • Receptors, Transforming Growth Factor beta / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Transforming Growth Factor beta1 / chemistry*
  • Transforming Growth Factor beta1 / metabolism

Substances

  • BMP4 protein, human
  • Bone Morphogenetic Protein 4
  • Receptors, Transforming Growth Factor beta
  • Recombinant Proteins
  • Transforming Growth Factor beta1