Label-free detection technology utilizes optics-based biosensors for the conversion of biological binding responses into signals without requiring fluorescent or any other label. It allows for the real-time monitoring of the changes that happen during analyte binding to an immobilized ligand on a biosensor surface, without individual assay components’ manipulation. The major benefit of the label-free technology is that it enables ligands to maintain their original conformation and biological activity.
This technology also mitigates the risk of the unwanted background signal that may be caused while binding labels non-specifically to other bioassay components, by eliminating labeled detection reagents’, such as fluorescently-labeled proteins and enzyme-conjugated antibodies, requirement. It is also advantageous while working with impure or complex sample material containing various components other than the analyte of interest.
Moreover, it facilitates researchers while performing analyses that are not possible with the conventional methods. Label-free monitoring enables affinity and kinetic analysis, including binding specificity evaluation, which are essentially aided by dynamic, real-time interaction analysis.
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Surface Plasmon Resonance
Surface plasmon resonance (SPR) is an optical technique for carrying out label-free detection. It provides accurate results without any chance of the normal functioning of the ligand or analyte being affected upon binding with the labels. It also saves cost, as it does not involve the use of expensive radioactive fluorescent labels and other reagents. In addition, SPR facilitates the detection of the changes in the dissolved material’s mass in the solvent layer close to the surface of the sensor, by measuring the change in SPR angle or refractive index.
Binding Kinetics
Kinetic binding refers to the phenomena of how quickly a compound binds to its target and how quickly it disassociates. It primarily measures the on-rate and off-rate.
Application of Kinetic Binding in Drug Discovery
The drug-induced conformational changes and binding kinetics to physiological target determine how a dose-response relationship is influenced by coupling efficiency. Binding kinetics provides support in evaluating the affinity of a drug to its target, and it can also affect the efficiency of the drug-response relationship by impacting the equilibrium state, thus resulting in improved efficacy with a longer duration of the response and differentiation of the therapeutic indications, along with higher tolerability.
Moreover, in the series of typical and atypical antipsychotic drugs, the measurements of the kinetic binding properties, along with correlation of the side effects, birthed a new model of binding that reflects that association rates are primarily responsible for causing side-effects, not the dissociation rates. It creates opportunities to build strategies to optimize kinetics at the D2 receptor and enhance the therapeutic profile of atypical antipsychotic drugs.
Therefore, the demand for label-free detection technology is rising due to the high efficiency and accurate results it provides without variations, as compared to conventional label-based technologies.
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