2. Academic Validation
  3. Quantitative, Real-Time Measurements of Intracellular Target Engagement Using Energy Transfer

Quantitative, Real-Time Measurements of Intracellular Target Engagement Using Energy Transfer

  • Methods Mol Biol. 2019;1888:45-71. doi: 10.1007/978-1-4939-8891-4_3.
Matthew B Robers 1,  James D Vasta 2,  Cesear R Corona 3,  Rachel Friedman Ohana 2,  Robin Hurst 2,  Manisha A Jhala 4,  Kenneth M Comess 4,  Keith V Wood 2
Affiliations

Affiliations

  • 1 Promega Corporation, Fitchburg, WI, USA. matt.robers@promega.com.
  • 2 Promega Corporation, Fitchburg, WI, USA.
  • 3 Promega Biosciences Incorporated, San Luis Obispo, CA, USA.
  • 4 AbbVie, Inc., North Chicago, IL, USA.
PMID: 30519940 DOI: 10.1007/978-1-4939-8891-4_3
Abstract

Intracellular target affinity and residence time are fundamental aspects of pharmacological mechanism (Lu and Tonge, Curr Opin Chem Biol 14:467-474, 2010). Although various robust biochemical approaches exist to measure these binding characteristics, analysis of compound binding with isolated targets may not accurately reflect engagement in the milieu of living cells. To realize the influence of cellular context, methods are needed that are capable of quantifying affinity and residence time in the presence of the intracellular factors that may impact target engagement. Bioluminescence resonance energy transfer (BRET) offers a solution for intracellular target engagement when quantitative metrics or kinetic analyses are required.

Keywords

BRET; Intracellular affinity; NanoBRET(TM); Residence time; Target engagement.

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