Flow-through particles for the high-performance liquid chromatographic separation of biomolecules: perfusion chromatography

This paper reports a new technique for reducing resistance to stagnant mobile phase mass transfer without sacrificing high adsorbent capacity or necessitating extremely high pressure operation. The technique involves the flow of liquid through a porous chromatographic particle, and has thus been termed "perfusion chromatography". This is accomplished with 6000-8000 A pores which transect the particle. Data from electron microscopy, column efficiency, frontal analysis and theoretical modelling all suggest that mobile phase will flow through these large pores. In this manner, solutes enter the interior of the particles through a combination of convective and diffusional transport, with convection dominating for Peclet numbers greater than one. The implications of flow through particles on bandspreading, resolution and dynamic loading capacity are examined. It is shown that the rate of solute transport is strongly coupled to mobile phase velocity such that bandspreading, resolution of proteins and dynamic loading capacity are unaffected by increases in mobile phase velocity up to several thousand centimeters per hour. The surface area of this very large-pore diameter material is enhanced by using a network of smaller, 500-1500 A interconnecting pores between the throughpores. Scanning electron micrographs show that the pore network is continuous and that no point in the matrix is more than 5000-10,000 A from a through-pore. As a consequence, diffusional path lengths are minimized and the large porous particles take on the transport characteristics of much smaller particles but with a fraction of the pressure drop. Capacity and resolution studies show that these materials bind and separate an amount of protein equivalent to that of conventional high-performance liquid chromatography as well as low performance agarose-based media at greater than 10-100 times higher mobile phase velocity with no loss in resolution.     

Multidimensional chromatography coupled with mass spectrometry for target-based screening

Summary The synthesis of structural analogs and the process of drug discovery have evolved dramatically through recent advances in solid-phase synthesis reagents and automated screening systems. As molecular diversity strategies emerge, the need for automated target-based selection of lead candidates becomes equally important. Multidimensional automated chromatographic techniques coupled to electrospray ionization mass spectrometry facilitate the selection process and provide maximum characterization information in a single screening run. The capture of tightly bound affinity leads by target biomolecules, followed by subsequent release and high-resolution separation with sensitive detection, significantly reduces the time required to identify and characterize lead compounds. This automated multidimensional chromatographic approach coupled with mass spectrometry, Selectronics, was used with several organic and natural libraries to demonstrate an automated target-based screening technique to select for high-affinity binders as potential lead compounds.Abbreviations ESI electrospray ionization - HPLC high-performance liquid chromatography - HTS high-throughput screening - ESI-TOF electrospray ionization time-of-flight - SAR structure-activity relationship     

Morphological Component Analysis and Inpainting on the Sphere: Application in Physics and Astrophysics

Morphological Component Analysis (MCA) is a new method which takes advantage of the sparse representation of structured data in large overcomplete dictionaries to separate features in the data based on the diversity of their morphology. It is an efficient technique in such problems as separating an image into texture and piecewise smooth parts or for inpainting applications. The MCA algorithm consists of an iterative alternating projection and thresholding scheme, using a successively decreasing threshold towards zero with each iteration. In this article, the MCA algorithm is extended to the analysis of spherical data maps as may occur in a number of areas such as geophysics, astrophysics or medical imaging. Practically, this extension is made possible thanks to the variety of recently developed transforms on the sphere including several multiscale transforms such as the undecimated isotropic wavelet transform on the sphere, the ridgelet and curvelet transforms on the sphere. An MCA-inpainting method is then directly extended to the case of spherical maps allowing us to treat problems where parts of the data are missing or corrupted. We demonstrate the usefulness of these new tools of spherical data analysis by focusing on a selection of challenging applications in physics and astrophysics.     

Simulations of kinetic electrostatic electron nonlinear (KEEN) waves with variable velocity resolution grids and high-order time-splitting

KEEN waves are non-stationary, nonlinear, self-organized asymptotic states in Vlasov plasmas. They lie outside the precepts of linear theory or perturbative analysis, unlike electron plasma waves or ion acoustic waves. Steady state, nonlinear constructs such as BGK modes also do not apply. The range in velocity that is strongly perturbed by KEEN waves depends on the amplitude and duration of the ponderomotive force generated by two crossing laser beams, for instance, used to drive them. Smaller amplitude drives manage to devolve into multiple highly-localized vorticlets, after the drive is turned off, and may eventually succeed to coalesce into KEEN waves. Fragmentation once the drive stops, and potential eventual remerger, is a hallmark of the weakly driven cases. A fully formed (more strongly driven) KEEN wave has one dominant vortical core. But it also involves fine scale complex dynamics due to shedding and merging of smaller vortical structures with the main one. Shedding and merging of vorticlets are involved in either case, but at different rates and with different relative importance. The narrow velocity range in which one must maintain sufficient resolution in the weakly driven cases, challenges fixed velocity grid numerical schemes. What is needed is the capability of resolving locally in velocity while maintaining a coarse grid outside the highly perturbed region of phase space. We here report on a new Semi-Lagrangian Vlasov-Poisson solver based on conservative non-uniform cubic splines in velocity that tackles this problem head on. An additional feature of our approach is the use of a new high-order time-splitting scheme which allows much longer simulations per computational effort. This is needed for low amplitude runs. There, global coherent structures take a long time to set up, such as KEEN waves, if they do so at all. The new code’s performance is compared to uniform grid simulations and the advantages are quantified. The birth pains associated with weakly driven KEEN waves are captured in these simulations. Canonical KEEN waves with ample drive are also treated using these advanced techniques. They will allow the efficient simulation of KEEN waves in multiple dimensions, which will be tackled next, as well as generalizations to Vlasov-Maxwell codes. These are essential for pursuing the impact of KEEN waves in high energy density plasmas and in inertial confinement fusion applications. More generally, one needs a fully-adaptive grid-in-phase-space method which could handle all small vorticlet dynamics whether pealing off or remerging. Such fully adaptive grids would have to be computed sparsely in order to be viable. This two-velocity grid method is a concrete and fruitful step in that direction.     

Direct experimental evidence for current-transfer mode operation of nested tungsten wire arrays at 16-19 MA

Nested tungsten wire arrays (20-mm on 12-mm diam.) are shown for the first time to operate in a current-transfer mode at 16-19 MA, even for azimuthal interwire gaps of 0.2 mm that are the smallest typically used for any array experiment. After current transfer, the inner wire array shows discrete wire ablation and implosion characteristics identical to that of a single array, such as axially nonuniform ablation, delayed acceleration, and trailing mass and current. The presence of trailing mass from the outer and the inner arrays may play a role in determining nested array performance.     

Tandem chirped quasi-phase-matching grating optical parametric amplifier design for simultaneous group delay and gain control

We present a broadband optical parametric amplifier design using tapered gain and tandem chirped quasi-phase-matching gratings to obtain flat gain and group-delay spectra suitable for applications such as ultrashort-pulse amplification and fiber-optic communication systems. Although a tapered-gain amplifier consisting of a single chirped grating can provide constant gain over a wide frequency range, it cannot be used to control the group delay across the spectrum. We propose controlling both the gain and the group delay profiles using a two-stage amplifier configuration, in which the idler of the first is used as the input signal of the second.