Dependence of the electroosmotic mobility on the applied electric field and its reproducibility in capillary electrophoresis.

Experimental results on the electroosmotic mobility in fused-silica capillaries are presented for different applied voltages and solutions of different pH. The electroosmotic mobility is shown to be dependent on the applied voltage and this dependence cannot be attributed to the temperature effects. Results of the electroosmotic mobility measurements are found to be dependent also on the electrophoresis unit they have been performed in. The explanation given and the relevant theory presented are based on the hypothesis that these effects are produced by a radial electric field inevitably existing in any electrophoresis unit. The concept of the limiting electrophoretic mobility, i.e. extrapolated to the zero applied voltage, is introduced in order to characterize the properties of the solution-wall interface. The slope of the electroosmotic mobility dependence on the applied voltage depends on the solution pH and the surroundings of the capillary. Theoretical estimations agree well with both experimentally found limiting mobilities and slopes. Long-term variations of the electroosmotic mobility are supposed to be related with the cation penetration into the capillary wall.

Use of taylor-aris dispersion for measurement of a solute diffusion coefficient in thin capillaries.

A method for the fast measurement of the diffusion coefficients of both small and large molecules in thin capillaries is reported. The method relies on Taylor-Aris dispersion theory and uses standard instrumentation for capillary zone electrophoresis. With this equipment, which consists of thin capillaries (50 to 100 micrometers in inner diameter), an injection system, detector ports, and computer data acquisition, a sample plug is pumped through the capillary at known velocity and the peak dispersion coefficient (D(*)) is measured. With the experimentally measured values of D(*) and flow velocity, and knowledge of the inner diameter of the capillary, the molecular diffusion coefficient (D) can be rapidly derived. For example, for ovalbumin a D value of 0.759 x 10(-6) square centimeter per second is found versus a tabulated value of 0.776 x 10(-6) square centimeter per second (error, 2 percent). For hemoglobin a D value of 0.676 x 10(-6) square centimeter per second is obtained versus a literature value of 0.690 x 10(-6) square centimeter per second (error, 1.5 percent).

Electroosmosis of polymer solutions in fused silica capillaries.

The classical von Smoluchowski equation predicts that the electroosmotic mobility generated by the wall zeta potential could be suppressed if the viscosity of the solution adjacent to the wall were extremely high. When performing runs in capillaries filled with polymer solutions (2% methyl cellulose solutions with viscosities of 25 cP), however, one consistently finds that the quenching of electroosmotic mobility is substantially less than predicted by the von Smoluchowski relationship. The electroosmotic flow is progressively suppressed with subsequent electrophoretic runs, suggesting a "dynamic coating" of the polymers onto the capillary wall. This progressive reduction of electroosmotic mobility tends to a plateau value which is still substantially higher than the value derived on the basis of the von Smoluchowski relationship. The following explanation is proposed: due to the very high shear rate in the electric double layer, the polymer molecules change their orientation and/or conformation, which lowers the fluid viscosity in this region. A scaling equation for electroosmotic mobility taking into account the non-Newtonian properties of polymer solutions is derived. It predicts electric field dependence of the electroosmotic mobility as the shear rate in the double layer is proportional to the electric field. Experimental measurements confirm the dependence of the electroosmotic mobility on the electric field.

Vaccination with SPf66, a chemically synthesised vaccine, against Plasmodium falciparum malaria in Colombia.

Preclinical and clinical studies have established the safety and immunogenicity of the chemically synthesised SPf66 malaria vaccine. The present study is a phase III randomised, double-blind, placebo-controlled, efficacy trial completed in La Tola, Colombia. 1548 volunteers over one year of age received three doses of either the vaccine (n = 738) or placebo (n = 810). Active and passive case detection methods were used to document clinical episodes of malaria among the study population. The follow-up period began one month after the third dose and lasted for one year. 168 and 297 episodes of Plasmodium falciparum malaria were documented in the SPf66 group and the placebo group, respectively; this corresponds to a crude protective efficacy of 38.8%. Incidence rates for first or only P falciparum malarial episodes were 22.3% per annum among the vaccinee group and 33.5% among the placebo group (RR = 1.5; 95% Cl 1.23, 1.84). Therefore, the protective efficacy of SPf66 against first or only episodes was 33.6% (95% Cl 18.8, 45.7), being highest in children aged 1-4 years (77%) and adults older than 45 years (67%). The estimated protective efficacy against second episodes was 50.5% (95% Cl 12.9-71.9). Our study shows that the chemically synthesised SPf66 malaria vaccine is safe, immunogenic, and protective against P falciparum malaria in semi-immune populations subject to natural challenge.

Steady-state heat transfer and thermal zone spreading in gel isoelectric focusing.

The zone spreading caused by a transverse pH profile due to a temperature gradient through the thickness of a gel slab is estimated. The temperature difference (delta T) between the upper and lower gel surfaces can be calculated as a function of the electric power dissipated in the gel and the gel dimensions. It is found that when delta T is only 1 degree C the zone spreading due to this thermal excursion is as high as 0.5 mm. Thus, an admissible delta T is found to be equal to 0.2 degrees C, since this corresponds to a thermal zone spreading of only 0.1 mm, i.e. the same order of magnitude as the spatial resolution of a laser scanner. A thermal zone spreading of 0.1 mm is compatible with a resolving power of 0.01 pH unit, the current limit of conventional isoelectric focusing in amphoteric buffers. A requirement for the thickness of a gel slab is formulated: e.g., at 40 W applied power (over a gel surface area of 25 x 11 cm), a thermal zone spreading limited to 0.1 mm can only be obtained with a gel thickness of approximately 170 microns.