The BIApages are a roundup of the most common methods and techniques to investigate Biomolecular Interaction Analysis. Central are the methods which determine when and how biomolecules interact. You can think of the association and dissociation but also thermodynamics and conformational changes in molecules and complexes. The techniques are not necessarily label- or immobilization free.
MicroScale Thermophoresis (MST) measures the motion of molecules along microscopic temperature gradients and detects changes in their hydration shell, charge or size.
When performing a MicroScale Thermophoresis (MST) experiment, a microscopic temperature gradient is induced by an infrared laser, and the directed movement of molecules is detected and quantified using either covalently attached dyes, fluorescent fusion proteins or intrinsic tryptophan fluorescence.
By combining the precision of fluorescence detection with the flexibility ...
Interaction kinetics describes the interaction between one or more components. After the interaction the components leave each other unchanged as opposed to enzyme kinetics. The interaction kinetics can be subdivided in three distinct phases.
Each of the three phases contains information over the interaction between ...
In SPR experiments, low responses are preferred over high responses. This is because high responses in general are affected by mass transport and non 1:1 interactions (6), (8). But how low can you go? This depends in the first place on the instrument of choice. The signal should be above the noise level of the baseline. The curves should be easily recognisable and replicates should overlay (4), (7).
To determine the noise level of the instrument, first equilibrate the system to minimize drift. Then inject flow buffer several times and observe the average response. ...
There are only a few really label- and immobilization free techniques which are able to measure biomolecular interactions. One of the most used is Isothermal Titration Calorimetry (ITC). The technique is simple to perform but needs quite some sample. Other techniques require that at least one component is immobilized to a supporting structure or sensor surface.
Of this category, Surface Plasmon Resonance (SPR) and Bio Layer interferometry (BLI) are the most common used. The sample consumption with SPR type instruments vary on the design but are moderate to low. BLI however uses very low quantities of the sample which can be recovered after the experiment. For Micro Scale Thermophoresis (MST) one of the interactants must be labelled (although there is a complete label-free instrument). This method uses very low amounts of sample and is relatively fast compared to SPR and BLI.
There are many techniques to explore bio molecules and their interactions. Some are both label- and immobilization free, other require labelling or immobilization. In the table is a round-up of techniques used in the study of bio molecules with an indication of the main information that can be obtained.
|SPR||Surface Plasmon Resonance||yes||no||kinetic|
|MST||Micro Scale Thermoforesis||no||yes/no*||equilibrium|
|ITC||Isothermal Titration Calorimetry||no||no||equilibrium|
|AUC||Analytical Ultra Centrifugation||no||no||equilibrium|
|Microbalance / Cantilever||yes||no|
|AFM||Atomic Force Microscopy||yes||no|
|High Performance capillary electrophoresis||no||?||equilibrium|
|LFII||Label-Free Intrinsic Imaging|
|NMR||Nuclear Magnetic Resonance||structural|
|FRET||Fluorescent Energy Transfer||yes||equilibrium|
|Size exclusion chromatography||no||no||size / dimer|
* Depending on the type of instrument.
Roundup of the websites and suppliers I found that do measurements in non invasive ways on non labelled molecules.
|MicroCal||VP-ITC system which uses differential scanning calorimetry.|
|Axela Biosensors||Diffractive optics technology with grating-based light diffraction and immobilized capture surfaces.|
|Q-Sense||The quartz crystal microbalance consists of a thin quartz disc with electrodes on both sides. The resonance frequency depends on the attached mass that is oscillating. A viscoelastic film will dampen the oscillation.|
|BioForce Nanosciences||Atomic-force-microscopy-based platform for nanoarray analysis.|
|DeltaDot||High performance capillary electrophoresis with label-free intrinsic imaging using algorithms to identify biomolecules.|