Biomolecular interactions kinetics

The interaction between c Fusarium moniliforme and PGIP from Phaseolus vulgaris L. was investigated using a biosensor technique based on sur ce plasmon resonance (BIAlite). This new analytical system provides information on the strength and the kinetics of biomolecular interactions. 

Optical methods are a perfect tool to characterize interaction processes between a sensitive chemical or bio polymer layer and analytes1. Time-resolved measurements of this interaction process provide kinetic and thermodynamic data. These types of sensors allow the monitoring of production processes, quantification of analytes in mixtures and many applications in the area of diagnostics, biomolecular interaction processes, DNAhybridization studies and evenprotein/protein interactions2,3. 

Fluorescent or other labels can, in some cases, interfere with the molecular interactions or cell biology of target receptors, thus resulting in false information [6]. Labels, particularly for small molecules, can be difficult to be optimized, thus introducing additional complexity and assay development time to the process. Label-free detection methods not only overcome the problem of steric hindrance of a label, but also enable kinetic measures of biomolecular interactions [46]. 

For the biochemist, the advance of new, sensitive temperature-measuring devices comes just at a time when large amounts of very pure proteins, nucleotides, DNA, RNA, lipids, etc., are becoming available. Thus, the ubiquitous nature of heat can be utilized, providing both the thermodynamic and kinetic data needed for any theory of biomolecular interaction. 

Concepcion J, Witte K, Wartchow C et al (2009) Label-free detection of biomolecular interactions using biolayer interferometry for kinetic characterization. Comb Chem High Throughput Screen 12 791-800... 

The mean-field effect of the environment can be included in biomolecular simulations simply by adding an expression for the solvation free energy of an instantaneous solute conformation to a given molecular mechanics force field [1]. Such an implicit solvent potential addresses the thermodynamic component of solute-solvent interactions. Kinetic and hydrodynamic properties may be reintroduced through the use of Langevin dynamics where coupling with a temperature bath is implemented through stochastic collisions and solvent friction [2,3,18]. 

Test and measurement sensors, pressure transducers, torque sensors, instrumentation, accelerometers, and displacement sensors An extensive array of instruments and systems for applications ranging from automotive R D, process and environmental monitoring, in vitro medical diagnostics Surface plasmon resonance biosensors for the determination of the affinity and kinetics of biomolecular interactions... 

KCE establishes a new paradigm separation methods can be used as comprehensive kinetic tools. The majority of previous attempts to utilize chromatography and electrophoresis for studying biomolecular interactions were limited to assuming equilibrium between interacting molecules. Not only does such an assumption limit applications to measuring equilibrium constants, but also this assumption is conceptually mistaken since separation disturbs equilibrium. Kinetics must be appreciated when separation methods are used for studies of noncovalent interactions. This appreciation can dramatically enrich analytical capabilities of the methods. 

Another important consideration is the working frequency. To characterize the sheer number of sensors in the same array in real time, both the switching circuit and measurement units need to work at high enough frequency. The speed needed eventually depends on the nature and kinetics of the biomolecular interaction to be measured. For example, considering a typical array with 1,000 elements to be measured in 1 second, the logic circuit to address the... 

SPR offers several advantages over conventional techniques such as fluorescence or enzyme-Unked immunosorbent assay. Here the analyte does not require any special characteristics or labels and can be detected directly the measurements can be performed in real time to collect kinetic data it is a versatile technique, capable of detecting analytes over a wide range of molecular weights and binding affinities. Because of its unique features, SPR has become a powerful tool for studying biomolecular interactions. 

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