Binding Affinity Measurement
Binding affinity under biologically relevant conditions
FIDA technology offers precise binding affinity measurements in solution, providing absolute data that accurately reflects the strength of molecular interactions under biologically relevant conditions. Whether working with proteins, small molecules, or larger molecular complexes, FIDA allows you to measure binding affinity (Kd) without the need for immobilization and with a range from weak milimolar bindings to strong picomolar bindings.
By delivering absolute Kd values, through direct observation of how the apparent size change as the concentration of the analyte is increased, FIDA eliminates the need for calibration curves or external standards. Its ability to measure binding directly in the sample’s native environment makes FIDA the ideal choice for researchers seeking to study molecular interactions in their natural native environment. This reduces potential distortions caused by traditional methods that require surface immobilization or other sample modifications.
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Why does FIDA Binding Affinity measurement move frontiers?
Absolute Affinity Data:
FIDA provides direct, absolute Kd values without needing indirect calculations or secondary standards, ensuring high accuracy and facilitating the usage of affinity measurements in AI models.
In-Solution Measurement:
All binding affinity measurements are performed directly in solution, preserving the natural state of the molecules and interactions, avoiding artifacts that can arise with immobilization techniques.
No Need for Labels:
Since FIDA measures the interactions in their native form, no labels or probes are required, reducing the risk of altering the molecules’ behavior. (Fida Neo comes with detecors of 280nm, 480nm & 640 nm)
Minimal Sample Requirement:
Only small amounts of sample are needed to measure binding affinity, conserving precious materials.
Versatility Across Sample Types:
Whether working with purified proteins, small molecules, or even complex, unpurified samples (fermentation media, patient samples, cell lysate etc.), FIDA delivers accurate binding data.
Multi-Parameter Efficiency:
FIDA not only measures binding affinity but also provides simultaneous data on other aspects of molecular interactions e.g. kinetics (read more here), quanitfication or aggregation.
Why use the hydrodynamic radius (Rh) to measure binding affinity?
FIDA measures binding affinity based on changes in the hydrodynamic radius (Rh) of molecules when they interact in solution. As molecules bind, their size in solution (Rh) changes, which FIDA tracks to determine binding events and calculate the dissociation constant (Kd). This approach provides direct, real-time insight into how molecules interact under natural conditions.
Preservation of Native States:
Since the binding events are monitored in solution, the molecules retain their natural form and function, leading to more biologically relevant data.
Accurate, Absolute Data:
The change of apparent Rh directly exposes the molecular interaction to measurement. This allows FIDA to deliver absolute Kd values, unlike other methods that provide relative or indirect data. This absolute readout makes a direct impact on research capacities, and fundamentally differentiates FIDA from other in-solution technologies which do not deliver direct/absolute measurements.
No surface immobilization
or sample modifications:
Traditional methods like Surface Plasmon Resonance (SPR) require immobilization, which can alter the natural binding dynamics. FIDA avoids this by measuring interactions in their native, solution-based environment.
Biologically Relevant Data:
By measuring in solution, FIDA captures interactions as they occur naturally, ensuring that the binding affinity data reflects real-world molecular behavior.
Enhanced Sensitivity:
The ability to track small changes in Rh provides sensitivity to weak interactions that might otherwise be missed by more traditional methods.
No Interference from
Surfaces or Labels:
The lack of immobilization or labeling reduces the risk of experimental artifacts, leading to more trustworthy and reproducible results.
Speed and Efficiency:
FIDA allows for binding affinity to be measured alongside other key parameters (like size), reducing the number of experiments required and increasing overall productivity.
How does FIDA relate to other methods?
Getting closer to native conditions thanks to in-solution measurements
The comparison table highlights several key advantages of FIDA for binding affinity measurements. Unlike Surface Plasmon Resonance (SPR), FIDA performs in-solution measurements without requiring surface immobilization, ensuring more accurate and biologically relevant results. FIDA also stands out for its ability to work with unpurified, complex samples, offering greater flexibility than techniques like ITC or Dynamic Light Scattering (DLS), which often require purified or large sample volumes.
Detecting both strong and weak interactions with one technology
In terms of sensitivity, FIDA is capable of detecting both strong and weak interactions, providing reliable data across a wide range of binding affinities, which many traditional methods struggle to capture. Additionally, FIDA’s capacity to deliver multiple parameters in one assay greatly improves experimental efficiency, saving time and reducing the need for multiple experiments. Overall, FIDA’s combination of low sample requirements, high sensitivity, and the ability to work with real-world, complex samples positions it as a highly efficient and versatile tool for researchers.
Learn more during an exploratory call with Fidabio team.
We are happy to answer all of your questions. Book an exploratory call to learn more about FIDA and the match between your personal needs and what we can deliver. The call is non-binding and free of any charges, so feel free to fill the form!