Biomedical subjects

T W Root

Publications and source records attributed to T W Root.

4 recordsLinked to original sources

PubMed2003

Adsorptive membrane chromatography for purification of plasmid DNA.

Adsorptive membranes were investigated for the downstream processing of plasmid DNA by quantifying both separation efficiencies and adsorption uptake with the anion-exchange membranes. Separation efficiencies of the 10-ml Mustang-Q were measured using pulses of 6.1-kilo base pair plasmid DNA and lysozyme tracers, and comparing the responses for both conventional and reverse-flow operation. The plasmid exhibited nearly 200 plates/cm, almost as high efficiency as the protein despite the large difference in size. This behavior contrasts strongly with typical behavior for spherical porous particle packings, which predicted large decreases in efficiency with increases in tracer size. Batch adsorption isotherms for the 6.1-kilo base pair plasmid on small sheets of anion-exchange membranes at various ionic strengths showed high capacities for very large biomolecules. The maximum binding capacity for the membrane unit was calculated as 10 mg plasmid/ml, an order of magnitude greater than typical values reported for porous beads.

Adsorption↗

PubMed2001

Characterizing the performance of industrial-scale columns.

The performance of a large commercial chromatographic column was investigated using a short pulse of a tracer and an extension of the reverse-flow technique. This technique permits separate determination of the unavoidable irreversible microscopic processes and the reversible effects of flow maldistribution, and allows for the separation of flow maldistribution in the flow distributors from flow maldistribution inside the packed bed. This analysis was performed on a 0.44 m Millipore IsoPak column using Cellufine GC 700, cellulosic-based media with an average particle diameter of 75 microm, for the stationary phase. The column efficiency was quantified by analysis of the effluent curve from a short pulse of a 5% aqueous acetone tracer. The study examined behavior of beds of different lengths (10-24 cm) and beds packed from different slurry concentrations (10-75% v/v). The slurry-packed columns were very uniform, and no significant macroscopic flow maldistribution was observed inside the column. The observed bed plate heights conformed to the predictions of available one-dimensional continuum models. Dispersion in the flow distributors was significant, corresponding to 15-25% of the intracolumn dispersion when the full 24 cm available bed length was used and a proportionally larger increase for shorter bed lengths. Thus, the headers are shown to produce a significant increase in the observed plate height.

Chromatography, Liquid↗

PubMed1998

Refining the scale-up of chromatographic separations.

The use of heavily loaded columns and complex processing conditions makes scale-up of chromatographic separations a non-trivial process. The wide ranges of process conditions that must be investigated demands that a large number of preliminary experiments must usually be made in small columns and laboratory-scale work stations. These preliminary data can be biased by improper column packing, poor distributors and dispersion in auxiliary apparatus, and it is important to understand these disturbing factors in detail. Moreover, it is precisely at this macroscopic level that our understanding of the chromatographic process is weakest, for large columns as well as small. This paper addresses three of these factors: Efficient elimination of peripheral effects and characterization of both header flow distribution and packing non-uniformity. This will be done using a variety of experimental and analytical approaches including nuclear magnetic resonance imaging, computational fluid dynamics and mass transfer, and careful experimentation.

Acetone↗

PubMed1997

Measuring column void volumes with NMR.

A novel method for measuring resin porosities and column void volumes with fluorine. NMR has not been developed. In situ measurements of the void volumes accessible to an array of fluorinated probe molecules are used to characterize the pore size distribution of the media. Application of this simple procedure is demonstrated for a commercially packed column and several bulk resins. The porosity distributions obtained by this technique are similar to those obtained by size exclusion chromatography. Unlike chromatographic tracer studies, however, this method does not require packed columns.

Magnetic Resonance Spectroscopy↗