Dual-channel OpenSPR-2 accelerates accurate detection of proteinA and IgG kinetics

OpenSPR's innovative patented LSPR technology, the next generation of surface plasmon resonance (SPR), can be used to analyze all types of interactions, including real-time label-free detection including protein-protein, protein-small molecules, antibody-antigen, protein-nucleic acid, protein-adapted Body, protein-lipid, protein-carbohydrate, etc. One of the main areas of research is the study of protein-protein binding kinetics, as it helps researchers understand the biological processes involved in millions of proteins. Many scientists are also applying techniques such as SPR to determine the kinetic constants of interactions between these proteins.

Why study protein-protein interactions?

Protein interactions are critical for understanding disease pathways, metabolism, signal transduction, membrane transport, virulence, and therapeutic targets. At any given time, in any given cell, and in any organism that maintains the basic processes of homeostasis, there are millions of protein interactions, so interprotein interactions are life-reactive The foundation is an insurmountable step in both drug research, disease mechanism research and basic biology research, and is the most crucial step in the analysis of the basis of biological activities.

Why use SPR to study protein-protein interactions?

When describing protein interactions associated with disease, signaling, or metabolic pathways, researchers typically rely on affinity-based qualitative techniques that provide only endpoint data such as Co-IP, ELISA, or yeast two-hybrid. Other technologies such as BLI, ITC or MST can provide quantitative data, but there are some disadvantages, including low repeatability, high sample consumption or the need for labeling.

In contrast, OpenSPR's next-generation SPR technology provides highly repeatable, label-free quantitative data with extremely low sample consumption. The researchers can be provided to determine the kon, koff and KD values ​​of the interactions in order to gain a deeper understanding of the entire process of binding.

Analysis of Protein-protein Interactions with OpenSPR

Below is an example of how to use the OpenSPR dual-channel instrument to analyze the binding kinetics of protein A interaction with IgG antibodies. Protein A is a virulence factor produced by S. aureus that promotes immune system evasion by tightly binding to the Fc region of an IgG antibody. Under the step-by-step guidance of the OpenSPR ligand fixation wizard software, the carboxyl sensor chip is cleaned and activated on both channels. In this experiment, channel 1 was used as a reference channel and channel 2 was fixed with protein A as a sensor channel.

After protein A is immobilized on channel 2, both channels are blocked to eliminate non-specific binding. Four concentrations of human IgG were prepared and injected into both channels and binding kinetics and affinities were calculated using a 1:1 binding model in the TraceDrawer analysis software. The figure and table below show how dual channel OpenSPR can easily obtain high quality and high reproducibility data by removing any unfixed and/or non-specifically bound analytes through the reference channel.

With the Tracedrawer software, you only need to set it in one step, select the reference curve, click OK, you can easily deduct the background; then the fitting obtains complete and accurate kinetic parameters: Ka, kd, KD.

Nicoya OpenSPR Molecular Interoperator Study the Advantages of Protein-protein Interactions

• Easy to operate - 1 hour to control;

• Complete kinetic parameter detection - Ka, Kd, ​​KD;

• High efficiency---loading, results, one-click analysis, results in 10min, complete kinetic test results in 1-2h;

• High precision – detection is not affected by temperature and buffer refractive index;

• No need for dedicated correction channels – negligible bulk effects;

• Real-time Co-IP detection - 2 hours out of test results, no need to transfer film and electrophoresis;