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Label-free methods using a resonant waveguide grating biosensor to determine GPCR signaling pathways

  • US 8,426,148 B2
  • Filed: 11/30/2007
  • Issued: 04/23/2013
  • Est. Priority Date: 10/06/2007
  • Status: Expired due to Fees
First Claim
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1. A label-free method for GPCR signaling pathway determination, the method comprising:

  • immobilizing live cells on a resonant waveguide grating (RWG) biosensor;

    contacting the immobilized live cells with a substance that elevates intracellular cAMP and activates protein kinase A in the live cells;

    contacting the substance-contacted live cells with a GPCR ligand;

    detecting and comparing the GPCR ligand-induced biosensor response of the live cells in the presence and absence of the substance; and

    determining the signalling pathways of the GPCR activated by the ligand, wherein the substance-induced inhibition of the GPCR ligand-induced response indicates that the GPCR signals via Gα

    s
    pathway, the substance-induced partial suppression of the GPCR ligand-induced response indicates that the GPCR signals via a Gα

    q
    pathway, and the substance-induced potentiation of the GPCR ligand-induced response indicates that the GPCR signals via a Gα

    i
    pathway, wherein the response is a dynamic mass redistribution (DMR) signal measured by the resonant waveguide biosensor, and the substance is a probe molecule to differentiate all classes of signalling mediated through the GPCR.

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