Multidimensional GPCR Profiling and Screening Using Impedance-Based Label-Free and Real-Time Assay

  • Ning Ke
  • Khanh Nguyen
  • Jeffery Irelan
  • Yama A. Abassi
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1272)

Abstract

GPCRs constitute one of the most sought-after targets in drug discovery because they are associated with conditions ranging from cardiovascular diseases, autoimmune diseases, inflammation, cancer, and diseases of the nervous system. Moreover, they are one of the most amenable targets for drug discovery because they can be modulated by small molecules, peptides, proteins, and antibodies. Therefore it may not come as a surprise that close to 40 % of the drugs that are currently on the market are targeting GPCRs. It has become evident that GPCR signaling is highly complex and may involve multiple or a subset of pathways depending on the interaction of a GPCR with an agonist or antagonist. It is imperative that any functional screening for GPCR activity integrates this complexity. In this assay protocol, we describe how the xCELLigence RTCA HT impedance-based platform which can be used for functional cell-based GPCR assays can be utilized for GPCR screening.

Key words

GPCR Impedance-based sensors High-throughput screening 

References

  1. 1.
    Eglen RM, Reisine T (2009) New insights into GPCR function: implications for HTS. Methods Mol Biol 552:1–13PubMedCrossRefGoogle Scholar
  2. 2.
    Kenakin TP (2009) Cellular assays as portals to seven-transmembrane receptor-based drug discovery. Nat Rev Drug Discov 8:617–626PubMedCrossRefGoogle Scholar
  3. 3.
    Yu N et al (2006) Real-time monitoring of morphological changes in living cells by electronic cell sensor arrays: an approach to study G protein-coupled receptors. Anal Chem 78:35–43PubMedCrossRefGoogle Scholar
  4. 4.
    Stallaert W et al (2012) Impedance responses reveal beta(2)-adrenergic receptor signaling pluridimensionality and allow classification of ligands with distinct signaling profiles. PLoS One 7:e29420PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Gibson SK, Gilman AG (2006) Gialpha and Gbeta subunits both define selectivity of G protein activation by alpha2-adrenergic receptors. Proc Natl Acad Sci U S A 103:212–217PubMedCentralPubMedCrossRefGoogle Scholar
  6. 6.
    Zhang J-H, Chung TDY, Oldenburg KR (1999) A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J Biomol Screen 4:67–73PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Ning Ke
    • 1
  • Khanh Nguyen
    • 1
  • Jeffery Irelan
    • 1
  • Yama A. Abassi
    • 1
  1. 1.ACEA BiosciencesSan DiegoUSA

Personalised recommendations