17 research outputs found
Single-molecule techniques in biophysics : a review of the progress in methods and applications
Single-molecule biophysics has transformed our understanding of the
fundamental molecular processes involved in living biological systems, but also
of the fascinating physics of life. Far more exotic than a collection of
exemplars of soft matter behaviour, active biological matter lives far from
thermal equilibrium, and typically covers multiple length scales from the
nanometre level of single molecules up several orders of magnitude to longer
length scales in emergent structures of cells, tissues and organisms.
Biological molecules are often characterized by an underlying instability, in
that multiple metastable free energy states exist which are separated by energy
levels of typically just a few multiples of the thermal energy scale of kBT,
where kB is the Boltzmann constant and T the absolute temperature, implying
complex, dynamic inter-conversion kinetics across this bumpy free energy
landscape in the relatively hot, wet environment of real, living biological
matter. The key utility of single-molecule biophysics lies in its ability to
probe the underlying heterogeneity of free energy states across a population of
molecules, which in general is too challenging for conventional ensemble level
approaches which measure mean average properties. Parallel developments in both
experimental and theoretical techniques have been key to the latest insights
and are enabling the development of highly-multiplexed, correlative techniques
to tackle previously intractable biological problems. Experimentally,
technological developments in the sensitivity and speed of biomolecular
detectors, the stability and efficiency of light sources, probes and
microfluidics, have enabled and driven the study of heterogeneous behaviours
both in vitro and in vivo that were previously undetectable by ensemble
methods..
AUTONOMOUS AND ENVIRONMENTALLY COMFORTABLE TYPE OF HOUSING FOR THE DEVELOPMENT OF ARCTIC
AUTONOMOUS AND ENVIRONMENTALLY COMFORTABLE TYPE OF HOUSING FOR THE DEVELOPMENT OF ARCTIC
Establishment and application of a new method for the determination of kinetic parameters by plug-plug kinetic capillary electrophoresis (ppKCE)
Sweeping Capillary Electrophoresis: A Non-Stopped-Flow Method for Measuring Bimolecular Rate Constant of Complex Formation between Protein and DNA
Peak-Shape Correction to Symmetry for Pressure-Driven Sample Injection in Capillary Electrophoresis
Evaluation of an in-capillary approach for performing quantitative cytochrome P450 activity studies
Online Enantioselective Capillary Electrophoretic Method for Screening Cytochrome P450 3A4 Inhibitors.
The market share of single-enantiomer drugs is steadily increasing. The pharmacodynamics and pharmacokinetics of individual enantiomers can differ considerably. Thus, their characteristics have to be addressed as early as possible in the development process of new pharmaceuticals. Capillary electrophoresis is a promising technique for enantioselective drug metabolism studies due to highly effective separations, minuscule consumption of sample and reagents, compatibility with a variety of detection techniques, high-throughput via automation, and the implementation of online procedures. An online method comprised of the diffusion-based mixing of cytochrome P450 3A4 with racemic ketamine, incubation of the enzyme reaction, separation of the reaction products S- and R-norketamine, and their quantification is presented in this chapter. Since diffusion is an inherent property of all molecules, the method enables the addition of virtually any compound to the reaction mixture without the need for additional optimization of the mixing conditions, and thus can be favorably used for the rapid screening of putative cytochrome P450 3A4 inhibitors