ProteOn™ System Introduction

Introduction ProteOn System Analysis Published Data FAQ Resources

Scientists from Creative Biolabs are willing to provide custom antibody affinity measurement using ProteOn system. The system is a surface plasmon resonance (SPR) biosensor platform that provides a label-free and real-time analysis of binding kinetics of molecular interactions. The key applications include:

Identification of biomolecular interaction.
Antibody screening and ranking.
Validation of drug targets.
Binding kinetic determination.
Epitope binning and epitope mapping.
Other dynimic assay of molecular interaction.

Introduction

Similar to Biacore, ProteOn system detects protein-protein interaction based on SPR technology, which is an extremely sensitive method for detecting molecular interactions by tracking the change of signal measured by sensor chips. Different from using ELISA which is an end-point assay, with ProteOn, you can obtain full kinetic parameters related to the following questions:

How specific is the interaction
How fast is the interaction (K_a)
How stable is the ligand-analyte complex (K_d)
How strong is the affinity (K_D)

Affinity Measurement and Kinetic Screening by ProteOn System

Compared to classic Biacore system, ProteOn has a unique 6×6 array to generate an analysis of up to 6 ligands with up to 6 analytes simultaneously. This technique significantly increases the throughput of SPR sensor and provides a cost-effective choice (money and time) for antibody affinity measurement and kinetic screening with the performance of screening/optimization/characterization without the need of regeneration as its most significant character.

Using ProteOn system, we can perform these analyses:

1×1 Full optimization, to determine optimal assay conditions.
6×1 Kinetic screening, to measure kinetics under different conditions in the same run for more robust data.
6×6 Single-point screening, to screen libraries for interactions of interest.
36×1 Target screening (unique to SPR technology), screen a single analyte against 36 different targets in a single injection.

ProteOn System

ProteOn system can present kinetic characteristics by immobilizing 6 ligands on the surface with 6 different concentrations of the same analyte flowing across to collect 6 full kinetics within one run. Furthermore, a multiplex screening with 6 totally different targets and analytes is possible to provide the highest throughput within the shortest time. This unique feature of ProteOn not only enables a detection of up to 36 separated interactions in a single run, but also allows in-line parallel referencing. These spots employing unmodified act as references (interspot reference) can correct refractive index change and nonspecific binding. The real-time injection references help to adjust the baseline drift resulted from the changes of the ligand surface and collect high-quality SPR data with low signal-to-noise ratio, as often seen in the case of small molecule analytes. Other references can remove the bulk effect, such as real-time double reference and channel reference.

In traditional kinetic measurement with SPR, regeneration steps are usually required after binding assay between each analyte. The ligand surface regeneration must completely remove the analytes while remain the immobilized ligands unchanged. However, in ProteOn system, the regeneration of the surface is not required since the interactions occur separately in the featured 6x6 array.

Other optional Antibody Affinity Measurement Services:

Published Data

Interactions of BSA with Anti-Cancer Compounds Using ProteOn System

Fig. 3 Sensorgrams for the interactions of BSA with the six test compounds. (Ling Zhang, 2016)

Here, the researchers used ProteOn system biosensors and molecular docking studies to determine the interaction of a group of anticancer compounds with bovine serum albumin (BSA). The results showed that six anticancer compounds could reversibly bind to the immobilized BSA. The sensing diagrams of the six compounds are in accordance with the interaction model as a whole, and the binding kinetics can be analyzed. However, there were significant differences in their affinity for BSA. Understanding the detailed affinity and kinetic information of these anticancer compounds can provide important insights for optimizing the interaction and activity of potential compounds in the process of drug development.

References

Abdiche Y N, Malashock D S, Pinkerton A, et al. Exploring blocking assays using Octet, ProteOn, and Biacore biosensors[J]. Analytical biochemistry, 2009, 386(2): 172-180.ProteOn XPR36, protein interaction array system
Zhang, Ling, et al. "Interactions of bovine serum albumin with anti-cancer compounds using a ProteOn XPR36 array biosensor and molecular docking." Molecules 21.12 (2016): 1706.

FAQ

What is the ProteOn system, and how does it facilitate the detection of ligand-analyte interactions?
The ProteOn system is a surface plasmon resonance (SPR) instrument designed to monitor real-time, label-free interactions between biomolecules. It allows for simultaneous analysis of multiple interactions, making it highly efficient for studying the binding kinetics and affinity of ligands and their respective analytes. This system uses a microfluidic chip to flow analytes over a sensor surface where ligands are immobilized, detecting interaction by measuring changes in the refractive index near the surface.
Can the ProteOn system be used for both high-affinity and low-affinity interactions?
ProteOn system is versatile and capable of analyzing a wide range of interactions, from high to low affinity. Its advanced technology allows researchers to adjust the flow rate and concentration of analytes, providing flexibility to optimize conditions for detecting both strong and weak interactions effectively.
How does the ProteOn system handle complex samples?
ProteOn system is designed to handle complex biological samples, including serum, plasma, and cell supernatants. It employs robust surface chemistry and advanced fluidics that minimize non-specific binding and maximize the accuracy of interaction measurements. This makes it suitable for use in various applications, including drug discovery, antibody characterization, and basic research in complex biological matrices.
What are the advantages of using the ProteOn system over other interaction detection methods?
ProteOn system offers several advantages, including the ability to perform parallel analyses, which significantly speeds up the process of characterizing multiple interactions at once. Additionally, its label-free detection eliminates the need for secondary reagents, reducing the risk of interference and preserving the native state of the biomolecules. This system also provides comprehensive kinetic data, which is crucial for a detailed understanding of the interaction dynamics.
What types of ligands can be studied using the ProteOn system?
ProteOn system is capable of studying a wide array of ligands including proteins, peptides, small molecules, nucleic acids, and even whole cells. Its versatile surface chemistry allows for the immobilization of different types of biomolecules, facilitating studies on a diverse range of biochemical and pharmacological interactions.
How does temperature control in the ProteOn system affect interaction analysis?
Temperature is a critical factor in biomolecular interactions, influencing the kinetics and stability of binding events. The ProteOn system includes precise temperature control, allowing researchers to conduct experiments at consistent temperatures or to explore the temperature dependence of interactions. This feature is crucial for accurately mimicking physiological conditions and for studying thermodynamics.
Can ProteOn system differentiate between specific and non-specific binding?
ProteOn system is equipped to distinguish between specific and non-specific binding through its advanced software and sensor technology. By analyzing the association and dissociation rates of molecules at different concentrations and under various conditions, the system can help identify and characterize non-specific interactions, ensuring that only specific binding events are considered in the final analysis.