Membrane chromatography: Protein purification from E. coli lysate using newly designed and commercial anion-exchange stationary phases

This contribution describes the purification of anthrax protective antigen (PA) protein from Escherichia coli lysate using bind-and-elute chromatography with newly designed weak anion-exchange membranes. Protein separation performance of the new AEX membrane adsorber was compared with the commercial Sartobind^® D membrane adsorber and HiTrap™ DEAE FF resin column under preparative scale conditions. Dynamic protein binding capacities of all three stationary phases were determined using breakthrough curve analysis. The AEX membrane showed higher binding capacities than the Sartobind^® D membrane at equivalent volumetric throughput and higher capacities than the HiTrap™ DEAE FF resin column at 15 times higher volumetric throughput. Anion-exchange chromatography was performed using all three stationary phases to purify PA protein. Quantitative SDS-PAGE analysis of effluent fractions showed that the purity of PA protein was higher for membrane adsorbers than the HiTrap™ DEAE FF resin column and was the same for the new AEX membrane and Sartobind^® D membrane adsorbers. The effects of E. coli lysate load volume and volumetric flow rate on PA protein separation resolution using the membrane adsorbers were minor, and the peak elution profile remained un-changed even under conditions where >75% of the total protein dynamic binding capacity of the membranes had been utilized. PA protein peak resolution was higher using pH-gradient elution than with ionic strength gradient elution. Overall, the results clearly demonstrate that membrane chromatography is a high-capacity, high-throughput, high-resolution separation technique, and that resolution in membrane chromatography can be higher than resin column chromatography under preparative conditions and at much higher volumetric throughput.     

Extension of the angular spectrum method to calculate pressure from a spherically curved acoustic source

The angular spectrum method is an accurate and computationally efficient method for modeling acoustic wave propagation. The use of the typical 2D fast Fourier transform algorithm makes this a fast technique but it requires that the source pressure (or velocity) be specified on a plane. Here the angular spectrum method is extended to calculate pressure from a spherical transducer-as used extensively in applications such as magnetic resonance-guided focused ultrasound surgery-to a plane. The approach, called the Ring-Bessel technique, decomposes the curved source into circular rings of increasing radii, each ring a different distance from the intermediate plane, and calculates the angular spectrum of each ring using a Fourier series. Each angular spectrum is then propagated to the intermediate plane where all the propagated angular spectra are summed to obtain the pressure on the plane; subsequent plane-to-plane propagation can be achieved using the traditional angular spectrum method. Since the Ring-Bessel calculations are carried out in the frequency domain, it reduces calculation times by a factor of approximately 24 compared to the Rayleigh-Sommerfeld method and about 82 compared to the Field II technique, while maintaining accuracies of better than 96% as judged by those methods for cases of both solid and phased-array transducers.     

Measuring the effect of demagnetization in stacks of gadolinium plates using the magnetocaloric effect

The effect of demagnetization in a stack of gadolinium plates is determined experimentally by using spatially resolved measurements of the adiabatic temperature change due to the magnetocaloric effect. The number of plates in the stack, the spacing between them and the position of the plate on which the temperature is measured are varied. The orientation of the magnetic field is also varied. The measurements are compared to a magnetostatic model previously described. The results show that the magnetocaloric effect, due to the change in the internal field, is sensitive to the stack configuration and the orientation of the applied field. This may have significant implications for the construction of a magnetic cooling device.     

Ultra-short pulse laser ablation of copper, silver and tungsten: experimental data and two-temperature model simulations

Experimental results of femtosecond laser ablation of the metals copper, silver and tungsten are compared to simulations based on the two-temperature model. The comparison provides new information about the laser-heating process: For the noble metals (Cu, Ag), the energy transport via ballistic electrons must be included, while this effect is negligible for a transition metal (W). The comparison provides values for the range of ballistic electrons in the noble metals. The model calculation is also employed to investigate the dependence of the threshold fluence and melting depth on pulse duration. It is observed that for pulses shorter than approximately 1 ps the threshold fluence and melting depth are independent of the pulse duration, while they increase as τ ^0.47 and τ ^0.51, respectively, for pulses longer than ∼40 ps, in good agreement with approximate analytical expressions predicting a ?{t}\sqrt{\tau} dependence.     

Local Rings of Embedding Codepth 3. Examples

A complete local ring of embedding codepth 3 has a minimal free resolution of length 3 over a regular local ring. Such resolutions carry a differential graded algebra structure, based on which one can classify local rings of embedding codepth 3. We give examples of algebra structures that have been conjectured not to occur.     

Non-intrusive measurements of convective heat transfer in smooth- and rough-wall microchannels: laminar flow

The convective heat transfer behavior of laminar flow through a smooth- and two rough-wall microchannels is investigated by performing non-intrusive and spatially resolved measurements of fluid temperature via two-color fluorescent thermometry under constant heat flux conditions at three of the four microchannel walls. Pressure-drop measurements reveal that the apparent friction factors for all surfaces agree well with established macroscale predictions for laminar flow through rectangular ducts with the onset of transition at Re > Re_cr = 1,800 for smooth-wall flow and deviation from laminar behavior at progressively lower Re with increasing surface roughness. The local Nu for smooth-wall flow agrees well with macroscale predictions in both the thermally developing and developed regimes. With increasing roughness, while an enhancement in local Nu is noted for flow in the thermally developing regime, no measurable influence is noted upon attainment of a thermally developed state. These observations are supported by the examination of temperature profiles across the microchannel at various axial positions and Re, which suggest that the thermal boundary layer may be regenerated locally by roughness in the thermal entrance region of the flow resulting in an increased axial distance (compared to smooth-wall behavior) at which thermally developed flow is attained in the presence of roughness. Finally, estimates of the bulk Nu indicate enhancement in convective heat transfer over the smooth-wall case for laminar flow at higher Re while the smooth-wall bulk Nu data are found to agree well with macroscale predictions.     

A Convenient Synthesis of N-Methoxy-N-Methylamides from Carboxylic Acids

Carboxylic acids can be converted to their corresponding N-methoxy-N-methylamides in high yields using 2-chloro-1-methylpyridinium iodide as the coupling agent. The reaction proceeds without racemization when chiral carboxylic acids are used as the starting material.     

Ammonia synthesis and decomposition on a Ru-based catalyst modeled by first-principles

A recently published first-principles model for the ammonia synthesis on an unpromoted Ru-based catalyst is extended to also describe ammonia decomposition. In addition, further analysis concerning trends in ammonia productivity, surface conditions during the reaction, and macro-properties, such as apparent activation energies and reaction orders are provided. All observed trends in activity are captured by the model and the absolute value of ammonia synthesis/decomposition productivity is predicted to within a factor of 1–100 depending on the experimental conditions. Moreover it is shown: (i) that small changes in the relative adsorption potential energies are sufficient to get a quantitative agreement between theory and experiment (Appendix A) and (ii) that it is possible to reproduce results from the first-principles model by a simple micro-kinetic model (Appendix B).