Literature

Feel free to peruse our literature, including brochures, posters, videos, and background info!

Brochures

Agile R100 Brochure

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What can Agile R100 do? How does the technology work? What are some examples of measurements from the tool? See this overview brochure for answers!

Posters

Agile R100: An Orthogonal Biosensing Tool for Structure Activity Relationship (SAR) Studies During Early Stage Drug Discovery

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Presented at the Drug Discovery Chemistry Conference in San Diego in 2017.
This poster displays a simple SAR study performed with Agile R100 using TNFα as the immobilized target and three inhibitor compounds as analyte. The activity and affinity of the inhibitor compounds are quantified with dose-response curves and kinetic binding data.

 

Direct Detection of Borrelia Burgdorferi Antigen Using Field Effect Biosensing (FEB) Technology

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Presented at the Centers for Disease Control and Prevention (CDC) in Atlanta in 2016.
This poster depicts the detection of various concentrations of lysed Borrelia burgdorferi cells with AGILE R100.

 

 

Agile Sensors Quantify Interactions in Challenging Samples for Drug Discovery

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Presented at the Drug Discovery Chemistry Conference in San Diego in 2016.
This poster shows results from measurements with quantity-limited samples (50 µg of total protein) and at fM concentration.

 

Novel Protein Binding Kinetics Measurements in Complex Biological Samples Using Agile

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Presented at the Experimental Biology Conference in San Diego in 2016.

This poster shows results from measurements directly from cell lysate, protein extract, growth media, tissue lysate, and undiluted plasma.

Videos

How to Use the Agile R100 Label-free Kinetic Binding Assay

Agile R100 provides association and dissociation rates, dose-response curves, and small molecule interactions for drug discovery. Leveraging Field Effect Biosensing (FEB) technology, easy-to-learn Agile R100 measures in challenging crude samples with minimal volume. Validate hits with a non-radioactive, non-optical, all-electronic method and achieve measurements never before possible!

Background Info

Solvent Correction: An Additional Reference Measurement for Biomolecular Interactions

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Organic solvents induce large sensor responses in optical systems that can mask the lower magnitude sensor responses obtained from low molecular weight analyte. While solvent correction on optical systems allows for the subtraction of artifacts in a biomolecular kinetics data set arising from organic solvents, it requires carefully prepared reference samples, time to prepare these reference samples, and a dedicated reference cell. Field Effect Biosensing (FEB) does not require solvent correction because standard reference measurements can be taken in line with experiment samples, and the electrical nature of the sensing mechanism means solvents minimally affect the sensor response, dramatically reducing the need for extra sample preparation and tool run time.

Blocking and Quenching to Reduce Nonspecific Interactions in High Quality Biosensors

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The sensitivity and specificity of a biosensor are directly related to the availability and functionality of immobilized capture molecules, but ambient compounds may also nonspecifically contribute to the sensor response, thereby decreasing the signal-to-noise ratio (SNR). Blocking and quenching steps are used in biosensors to reduce the contributions from background compounds that interfere with the measurement signal. Several aspects of biosensor functionalization can be optimized to drive down the effects of nonspecific interactions and increase the SNR for targeted detection, critical to sensing low concentrations of target analyte with low signal. This technical note demonstrates effective blocking and quenching techniques with Agile R100 biosensor chips to deliver sensitive and specific sensor responses with high SNR.

User Manuals

Agile R100 User Manual

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Background detail about Field Effect Biosensing (FEB) technology and the Agile R100 system, experimental design, software setup, data acquisition, and data analysis.


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Comparison Studies

Agile R100 versus MicroCal iTC200 Kinetic Binding Data

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Agile R100 is a graphene biosensor with unprecedented functionality, providing label-free, real-time kinetic binding data. As a novel technology, Agile R100 is compared to MicroCal iTC200 to demonstrate its benefits compared to a standard kinetic binding analysis tool, isothermal titration calorimetry (ITC). In this study, kinetic binding data between the two instruments is compared by measuring Rho guanosine triphosphate hydrolase enzyme (GTPase) interacting with GTPase activating protein (GAP). The association and dissociation binding rate (ka and kd, respectively) and dissociation constants (KD) are reported. Agile R100 has a precise binding affinity measurement (KD = 1.35 ± 0.06 µM), comparable to MicroCal iTC200 (KD = 2.7 ± 0.3 µM), while also obtaining ka and kd values. Additionally, Agile R100 uses up to 30 million times less sample material and up to 30 times less sample volume and can perform 2.5 times more measurements in a given workday compared to MicroCal iTC200.

Application Notes

Detecting Interactions of Small Molecules in DMSO Using Agile R100

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Small molecules are beneficial as drug therapeutics but often have low solubility in aqueous solutions. Solvents such as dimethyl sulfoxide (DMSO) are necessary additives for small molecule solubility, but these solvents can interfere with optical biosensing platforms. In this technical note, Agile R100 successfully detects the interaction between the cytokine tumor necrosis factor alpha (TNFα) and the small molecule inhibitor SPD304 in 0%, 1%, 3%, and 10% DMSO. Because Agile R100 uses an electrical technique rather than an optical one, DMSO in buffer is compatible with the instrument and does not require solvent correction. Additionally, high concentrations of SPD304 (100 μM) were detected on Agile R100. Importantly, Agile R100 detects the association binding rate (ka) and the dissociation binding rate (kd) and can calculate the dissociation constant (KD) of the TNFα and SPD304 interaction in buffer with DMSO. Agile R100’s KD result (14.5 ± 1.8 μM in phosphate buffer saline [PBS] with 10% DMSO) is similar to a previously reported KD value (7.3 ± 0.5 μM in citrate phosphate buffer with 10% DMSO).

Affinity Ranking of SPD304, Evans Blue, and Trypan Blue to TNFα Using Agile R100

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Affinity ranking is critical to optimizing the relationship between a target protein and a potential drug compound during the hit-to-lead optimization phase of drug discovery. In this application note, Agile R100 characterizes the activity of the target protein TNFα interacting with the small molecule drug compounds, SPD304, Evans Blue, and Trypan Blue in order to rank order by affinity.

Technical Notes

Kinetic Binding Analysis on Agile R100

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Biosensors are used in a variety of fields to characterize biomolecular interactions in real-time. The results of these interaction studies inform the development of the next generation of diagnostics and therapeutics and provide insights into the mechanisms by which pathogens evolve and infect. Label-free assays are particularly useful in these endeavors because they eliminate both the chance of the added label interfering with the native binding chemistry and the additional chemistry steps to attach the label to the molecule under investigation. This technical note provides an overview of the kinetic binding data collected by Agile R100.

Reproducible Detection of Small Molecule Interactions Using Agile R100

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In this technical note, Agile R100 detects the interaction between the cytokine tumor necrosis factor alpha (TNFα) and the small molecule inhibitor SPD304 with high reproducibility between independent replicate measurements. Agile R100 detects the association binding rate (ka) and the dissociation binding rate (kd) and calculates the dissociation constant (KD) of the TNFα and SPD304 interaction with minimal deviation between independent replicate measurements (CV < 20%).

How Nonspecific Binding is Prevented on Agile R100

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To prevent nonspecific sensor responses during the experimental design, several different sensor functionalization chemistries are investigated on Agile R100, an assay that leverages Field Effect Biosensing (FEB) technology. The optimized sensor functionalization chemistry is used to demonstrate the biosensing capabilities of Agile R100 in complex media without the need for additional blocking steps, where Agile R100 demonstrates no loss of sensitivity to its target analyte in simulated serum compared to simple buffer.