Technology

FIDA stands for Flow Induced Dispersion Analysis. It is a first-principle based, in-solution technology, which provides a precise, quantitative, and highly robust method to determine how changes to an analyte affect the analyte. The changes that may be assessed are numerous ranging from adding a binding partner to changes in the environments including ionic strength, pH, buffers and different crude matrixes, temperature etc. The typical readout is hydrodynamic radi which provides information about quantity, affinity, sample integrity, changes in oligomeric state, aggregation, stoichiometry etc. Press to keep reading...

FIDA is based on 2 principles: Taylor Dispersion Analysis & Stokes-Einstein Equation. Utilising Taylor Dispersion Analysis, incl. the Stokes-Einstein Equation, FIDA in a very accurate way detects the change in apparent diffusivity that occurs when an analyte interacts with a binding partner or the environment changes, which causes a change in the hydrodynamic radius readout. Press to keep reading.

In-solution studies increase data reliability, due to their ability to mimic the natural physiological conditions in which proteins and particles function. Proteins and particles are sensitive structures. Their functions are often impacted by environmental changes, by labelling or by immobilization to a surface structure and functionality of the protein. All-in-all, in-solution studies increase data reliability, due to their ability to mimic the natural physiological conditions in which proteins and particles function. In solution approach allows to better understand the dynamic molecular interactions, conformational changes, and biochemical reactions, leading to more accurate insights into their behavior and enabling the development of more targeted and effective applications in various fields, including drug discovery, diagnostics, and biotechnology.

FIDA requires a fluorescent signal coming from your sample that can be detected by one of our detectors. It can either be emitting from a label or from the molecule itself, i.e. label-free. The signal coming from your sample that can be detected by one of our detectors: 480, 550, 640, 280. FIDA uses a LED excitation and amplifies the emission signal by PMT. Therefore, FIDA does not use any laser to get a signal, and the sample is not heated. Note that FIDA delivers two orthogonal readouts: absolute size (hydrodynamic radius) and fluorescence.

The main read-outs with FIDA are: - The Hydrodynamic radius (size in nm) - The Relative Fluorescence Unit (RFU)

Hydrodynamic radius (Rh), is a parameter used to describe the size of a particle or molecule in a fluid, taking into account its shape, mass, and interactions with the surrounding fluid molecules. The hydrodynamic radius is defined as the radius of a hypothetical sphere that diffuses at the same rate as the particle or molecule in question, under the same conditions (Einstein, 1905).

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FIDA can detect size changes smaller than 5%. In many cases you may detect changes down to an angstrom.

Molecular size and molecular weight are the two most used properties to characterise molecules. Molecular size can be approximated with molecular weight. Molecular size refers to the geometry of the molecule and is reported as the hydrodynamic radius (Rh). Rh is the absolute size, in nanometers, of the molecule in solution. On the other hand, molecular weight is calculated as the sum of the atomic weights of all the atoms in a molecule and is reported in kilodaltons (kDa).In general, larger sized molecules tend to have higher molecular weight than smaller molecules, but the relationship between molecular size and weight is not always straightforward. This is especially true when considering different conformational states of proteins, such as globular versus unfolded, as the complexity of measurements increases in this case. Press to keep reading...

Measuring binding affinity is achieved through Flow Induced Dispersion Analysis (FIDA) by quantifying changes in the hydrodynamic radius (Rh). As biomolecules bind in a solution, their diffusivity decreases, leading to slower diffusion and an extended dispersion profile. FIDA software, applying the Stokes-Einstein equation, precisely determines the increase in size (Rh) as complexes start to form. By titrating the molecule of interest with its binding partners in a straightforward FIDA experiment, binding affinity (Kd) can be accurately assessed based solely on the hydrodynamic radius and the Stokes-Einstein equation.

No, there is no buffer restriction with FIDA. Hurra! Crude matrix like serum, plasma, fermentation media, cell lysate works perfectly fine on FIDA, which increases clinical relevance of the studies made. On top of that, as the viscosity is measured and compensate at every measurement, high percentage of viscous additive (glycerol, DMSO, PEG…) can be added to your buffer prior a FIDA measurement.

No. Contrary to many other technologies, FIDA provides full buffer flexibility, incl. PEG, Glycerol, DMSO media etc. As the viscosity is measured and compensated at every measurement, a high percentage of viscous additives can be added to your buffer prior to a FIDA measurement.

Yes it is. As the viscosity is measured and compensate at every measurement, high percentage of viscous additive (glycerol, DMSO, PEG…) can be added to your buffer prior to a FIDA measurement.

No, FIDA provides precise viscosity measurements and any change in viscosity will be accommodated for in the FIDA data analysis. The Fida 1 can also be used as an advanced, highly accurate viscosity meter.

No, there is none.

It takes from app. 30 min. depending on the assay type.

Yes, the 96-well plates from FIDA are compatible with pipetting robots

Different binding models are used in the binding curve module in the Fida data analysis software. In general, the binding models are derived assuming that the apparent diffusivity of the indicator is a weighted average of the fractions bound and unbound of the indicator. The exported data can then be treated in generic spreadsheet-based analysis software and custom graphics can be designed for presentations and publications. Press to read more and see calculations.

Yes, Fida Neo can be used to measure kinetics (association kon and dissociation koff rates). It does so in solution, and with no buffer, temperature, detergent or ionic strength constraints.