Experimental observation of light-induced solitary waves of analyte bands in capillary electrophoresis.

The phenomenon of electrophoresis in free solution has been studied theoretically down to the molecular level for decades. In addition, intermolecular photo-induced proton transfer reactions, which occur in a wide class of molecules (phenols and aminoarenes) as well as proteins (green fluorescent protein), were also studied extensively. However, the study of the effect of light-induced electrophoretic mobility changes of the analytes in electrophoresis was begun only recently. In the present work, capillary zone electrophoresis was chosen as the environment to measure the magnitude of these electrophoretic mobility shifts induced by light. Background electrolytes (running electrolytes) with high refractive indices were developed, allowing the capillary to work like an optical fiber. The experimental conditions for obtaining stable coupling and guided laser light along the liquid core are discussed. Experimental evidence of band compression is observed, leading to a solitary wave behavior of the analyte band (2-naphthol). These solitary waves result from competition between thermal diffusion (dispersion mechanism) and a nonlinear (band compression) effect due to the combined electrophoresis phenomenon and absorption of guided light by the molecules of the band (which are subjected to a "reversible intermolecular proton transfer reaction" as one of their decay routes). The possibilities of applying this effect to different methods and techniques are also discussed.

Trapping electrophoresis and ratchets: a theoretical study for DNA-protein complexes.

Recently, Griess and Serwer (1998. Biophys. J. 74:A71) showed that it was possible to use trapping electrophoresis and unbiased but asymmetrical electric field pulses to build a correlation ratchet that would allow the efficient separation of naked DNAs from identical DNAs that form a complex with a bulky object such as a protein. Here we present a theoretical investigation of this novel macromolecular separation process. We start by looking at the general features of this electrophoretic ratchet mechanism in the zero-frequency limit. We then examine the effects of finite frequencies on velocity and diffusion. Finally, we use the biased reptation model and computer simulations to understand the band-broadening processes. Our study establishes the main experimental regimes that can provide good resolution for specific applications.

Recent developments in DNA electrophoretic separations.

DNA electrophoresis is now a fairly mature technology. Nevertheless, as we approach the 21st century, new ideas are frequently suggested that could lead to a revolution for DNA sequencing and mapping. Here, we review some of the novel concepts that have been studied since ca. 1990. Our review focuses on new separation mechanisms, new sieving matrices and recent conceptual advances.

Theory of solitary waves in electrophoresis.

A theoretical study about the generation of solitary waves (SW) in electrophoresis and capillary electrophoresis is performed. Two models that use velocity terms in Fick's first law and absorption and decaying terms in Fick's second law are presented. These models give the time evolution of the band or zone shape in electrophoresis and capillary electrophoresis in the presence of electromagnetic radiation. In particular, the effect of electrophoretic mobility changes of the analytes due to radiation excitation is included in the present models. The analytes are represented either by three-level or four-level systems. It is shown that the resulting system of coupled nonlinear partial differential equations governing the spatial motion of these bands over time exhibits SW solutions for some values of the equation parameters. We analyze the conditions in which these SWs, which propagate with constant velocity, constant area, constant standard deviation, and without change of form, are generated. The results of the present models are compared with those from a previous two-level model (Phys. Rev. Lett. 1995, 75, 1210-1213). The velocities of these SWs are calculated analytically. The time evolution of their standard deviations is shown. The numerical integration of a two-component electrophoresis run shows higher resolutions under some conditions. The expected practical difficulties which may be encountered when observing this phenomenon are discussed. Some practical difficulties that are likely to limit its useful application in electrophoresis and capillary electrophoresis are mentioned.

Capillary zone electrophoresis separation of kinins using a novel laser fluorescence detector.

1. Bradykinin, Lys-bradykinin, Met-Lys-bradykinin, des-Arg9-bradykinin and des-Arg1-bradykinin were separated by capillary zone electrophoresis in an apparatus constructed in our laboratory which utilizes a novel N2 pulsed laser-induced fluorescence detector. 2. Detection limits of 1.2 fmol for fluorescamine-derivatized bradykinin and 90 attomol for O-phthaldialdehyde-derivatized bradykinin were achieved. 3. This powerful analytical tool is described and its successful application to the measurements of bradykinin after enzymatic release from blood is documented.

Capillary zone electrophoresis separation of kinins using a novel laser fluorescence detector

1. Bradykinin, Lys-bradykinin, Met-Lys-bradykinin, des-Arg9- bradykinin and des-Arg1- bradykinin were separated by capillary zone electrophoresis in an apparatus constructed in our laboratory which utilizes a novel N2 pulsed laser-induced fluorescence detector. 2. Detection limits of 1.2 fmol for fluorescamine-derivatized bradykinin and 90 attomol for O-phthaldiadehyde-derivatized bradykinin were achieved. 3. This powerful analytical tool is described and its successful application to the measurements of bradykinin after enzymatic release from blood is documented