G-protein-coupled receptors (GPCRs) Analysis

Agile R100 is a label-free personal assay system that makes finicky GPCRs easy to study. Need to use detergents to solubilize your GPCR, or low temperatures to stabilize it? Having difficulty expressing and purifying your membrane protein, leaving you just a tiny bit of sample? Not a problem. Agile R100 uses a proprietary electrical technique that’s revolutionizing GPCR characterization capabilities, giving you the data you need for this critical drug target.

Why is GPCR characterization important?

Drug discovery is a long and expensive process costing an average of $1.5 billion, with a large amount of the expense due to candidate attrition in the later clinical stages. GPCRs are one of the most significant therapeutic target classes with 40-50% of all modern drugs acting on GPCRs in areas such as cancer, immune and inflammatory disorders, and neurological and metabolic diseases. If valuable kinetic characterization can be gained earlier in the drug discovery process, better decisions can be made in compound selection, potentially increasing the success rate of candidates and reducing expense and time to market.

Why is it difficult to develop assays to study GPCRs?

While GPCRs are crucial drug targets for therapeutic intervention, this target class is notoriously hard to analyze due to three main reasons: The difficulty of expressing and purifying GPCRs in functional form, the instability of the protein when solubilized, and the delicate balancing act required to develop assays that can successfully measure solubilized GPCRs.

How does Agile R100 overcome these issues?

Agile R100 is built with proprietary Field Effect Biosensing (FEB) technology, a breakthrough electrical technique. Advantages of the system that enable successful GPCR characterization include: low target density requirements, enabling the study of even a tiny amount of purified GPCR, a non-microfluidic format that allows the target to be applied directly to the biosensor surface, reducing protein degradation, temperature versatility to keep GPCRs within their required stability range, and the ability to sense in complex samples containing solubilizing detergents.

This means you can now study GPCRs in conditions that are more natural to the protein: in low concentrations with a non-flow assay that minimizes shear stress. The entire immobilization and measurement process can be done in under an hour, perfect for quickly-destabilizing proteins like GPCRs. Agile R100’s ability to sense in complex samples lets you measure in the optimal conditions for your protein without spending time in development to modify for the limitations of your platform.

Agile R100 features:

  • Detection in complex samples including solubilizing detergents without the need for time-consuming, error-prone reference subtraction.
  • Non-microfluidic format that applies the sample directly to the biosensor surface, promoting protein stability
  • Low target density requirements, letting you study even a small amount of purified GPCR
  • Temperature versatility. Agile R100 can be placed on a bed of ice, with real-time temperature monitoring built into Agile biosensor chips to ensure your GPCR is kept at a stable temperature.


GPCR chemokine receptor (CR) and Compound A

label-free Agile R100 kinetic characterization

In the figure above, a full dose-response curve for the interaction of GPCR CR and Compound A is shown, generated with NTA Agile biosensors. The Kfrom this methodology was 68.5 ± 17.7 µM, within a factor of 1.4 from the KD determined using single-concentration kinetics. With single-concentration kinetics highly indicative of full dose-response curve results, researchers can reduce the number of total measurements to save time and resources. For further details, see the Brite Paper in Downloadable Literature.