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Surface Plasmon Resonance (SPR) Analysis
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A Powerful Technique for Studying Molecular Interactions
By measuring the adsorption of molecules on a surface, surface plasmon resonance (SPR) can provide detailed information about binding events without the need for sample labeling.
Applications
- Binding Affinity: Determine the strength of the interaction between molecules, such as ligand-receptor or antibody-antigen interactions.
- Kinetics of Interaction: Measure the rates of association and dissociation, giving you insights into how quickly molecules bind and release.
- Concentration Analysis: Understand the concentration of analytes in a solution based on their binding response.
- Concentration Analysis: Understand the concentration of analytes in a solution based on their binding response.
What SPR Can Tell You
Surface Plasmon Resonance (SPR) provides real-time insights into molecular interactions by measuring changes in the refractive index on a sensor surface as molecules bind to it. Unlike mass spectrometry, which measures molecular mass, SPR focuses on the dynamics of molecular adsorption, revealing binding kinetics, affinity, specificity, and concentration. This technique is essential for understanding protein-ligand interactions, antibody-antigen binding, and other molecular phenomena.
Key Insights from SPR:
- Binding Kinetics: Rates of association and dissociation.
- Binding Affinity: Strength of interactions (KD value).
- Specificity: Selectivity of binding interactions.
- Concentration: Quantification of bound molecules.
- Interaction Mechanisms: Mechanisms and conformational changes upon binding.
Fig: An example SPR sensorgram showing stages of a typical cycle.
By varying the concentration of the analyte being measured accurate affinity measurements can be determined from multiple sensorgrams.
Fig: Example binding affinity measurement by taking measurements at different concentrations.
Using multiple sensorgrams, an accurate dissociation constant (KD) can be calculated for a given reaction.
How to Interpret SPR
Understanding SPR data involves analyzing the sensorgram, a real-time plot of the response signal versus time. Here’s how to interpret the key components:
- Association Phase: This phase shows the binding of molecules to the sensor surface. The rate and extent of binding provide insights into the binding affinity.
- Steady-State: This plateau phase indicates that the binding process has reached equilibrium, which helps in determining the equilibrium constant (Kd).
- Dissociation Phase: When the analyte is removed, this phase shows how quickly the molecules dissociate, revealing the stability of the interaction.
CovalX SPR Services
Our SPR analysis uses state-of-the-art equipment to detect minute changes in the refractive index near the sensor surface, ensuring high sensitivity and accuracy. This method is crucial for applications such as drug development, where understanding the interactions between potential therapeutics and their targets is essential.
Timeline, Sample Requirements and Resolution
- Sample Consumption: 75ug of each binding protein.
- Cost: CovalX offers upfront competitive pricing on a per-project basis.
- Affinity range: fM to mM binding
- Dissociation rate constant (kd): 10-5 to 1 s-1
- Sample type: All antibody formats accepted