HOW IT WORKS
AGILE R100 is a breakthrough electrical assay built with proprietary Field Effect Biosensing (FEB) technology that provides real-time label-free kinetic binding, affinity, and concentration data. At the heart of AGILE R100 is a biosensor chip built with graphene, a unique nanomaterial that delivers highly-sensitive biophysical characterization of small molecules ≥1 Da in complex media such as DMSO. A baseline read is taken with AGILE R100 prior to immobilizing the target. As the target is immobilized to the surface of the AGILE biosensor chip, the change can be seen in the built-in software’s sensorgram. As free-in-solution analyte is introduced, the association between target and analyte can be viewed in the real-time sensorgram as binding occurs. When dissociation occurs, the response decreases, which is also shown in the sensorgram. See the tabs in the below table for step-by-step details!
Measure in buffer
The surface of the AGILE biosensor chip is made of graphene, platinum, and glass, inert materials that do not react with your biology. A current is sent through the biosensor chip and surrounding solution to monitor for changes in conductance. When performed with buffer, this is the initial baseline data in the sensorgram.
Immobilize target to biosensor chip surface
As the target is immobilized, it changes how the current applied to the solution affects the graphene. Changes in any exposed charge groups or to the configuration of the target alters the conductance, and the change in conductance is captured in the output data.
Capture the binding interaction
When analyte binds to the immobilized target, it triggers a change in conductance which is captured and read by the AGILE R100 platform. The ability to measure this change effectively converts biological events (ex: target–analyte interactions) into sensitive electrical signals.
Dissociation of the analyte from the target will elicit the opposite response from association. As dissociation occurs, the immobilized target typically returns back to its starting structure, reversing the changes that led to the sensor response during association. This dissociation rate is also captured in real time.
View real-time data
AGILE R100 measures and records quantifiable changes in conductance in real time, providing accurate and specific kinetics, affinity, and concentration data.
HOW AGILE R100 SUPPORTS NEW CAPABILITIES
HIGH SENSITIVITY – AGILE biosensor chips are made with the nanomaterial graphene, which gives the platform its unprecedented 11 logs of dynamic range. Every atom of graphene is exposed to the sample, making AGILE R100 extremely sensitive to changes in conductance caused by binding interactions. This enables measurement even with minute amounts of material, small volumes and concentrations, and weak interactions.
SMALL VOLUMES – AGILE R100’s standard open-pipetting liquid handling design requires only 10 uL to measure. There is no need to coat the graphene sensor surface with excess protein; only a few thousand proteins are needed on each sensor for functionality. This enables functionalization using nM concentrations, with low volumes of material. Visit Data for example experiments using small sample sizes.
PORTABLE – AGILE R100 uses only electronic components which are small and cost effective. This is possible because AGILE biosensors are made with graphene, a material with game-changing electronic and chemical properties. The ability of graphene to continue functioning as a viable electronic material even when exposed to harsh buffers avoids the use of specialty optical, thermal, or nuclear components, reducing both size and cost.
In comparison, SPR requires expensive and bulky optical equipment. Isothermal Titration Calorimetry (ITC) requires a level of thermal insulation that again adds to the bulk and cost of the overall system, and Nuclear Magnetic Resonance (NMR) requires cryogenically cooled magnets.
RAPID MEASUREMENTS – AGILE R100 saves time in multiple ways. The open-pipetting design enables measurement within minutes of sample prep. Automated analysis software enables on-the-fly assay modification, as changes can be made to protocol during the experiment as data are reviewed. Because AGILE R100 is based on Field Effect Biosensing, an electrical method, the system can sense in optically dense sample. AGILE R100 has measured in human plasma, tissue lysate. Learn more in this poster titled AGILE Sensors Quantify Interactions in Challenging Samples for Drug Discovery, presented in 2016.
SMALL MOLECULE DETECTION – The Field Effect Biosensing (FEB) method on which AGILE R100 is based is fundamentally different from optical techniques. Optical tools such as SPR and BLI systems measure shifts in light caused by changes in mass, which is effective for large molecules. SPR and BLI struggle to measure interactions <1000 Da without solvent correction and additional calculations.
In contrast, FEB is an electrical technique, not a mass-based method. The size of the molecule does not impact the change in conductance experienced when an interaction occurs, enabling sensitive detection of biomolecules ≥1 Da. This makes FEB an excellent orthogonal technique for small molecule and fragment characterization and validation.
SENSE IN HIGH CONCENTRATIONS – Small molecule measurements often require high concentrations because interactions have kDs in the micromolar to millimolar range. For measurements using such high concentrations of small molecules, it is often necessary to include DMSO in the solution to maintain a known concentration and prevent precipitation.
The addition of DMSO has a large effect on the optical properties of a solution, and a 1% difference in DMSO concentration will lead to an optical sensor response ten times larger than the biochemical response, drowning out the interaction with background noise and requiring additional solvent correction measurements. AGILE is based on Field Effect Biosensing (FEB), an electrical technique, not an optical one, so there are no issues caused by optical limitations. Learn more about Detecting Small Molecule Interactions in DMSO Using AGILE R100.