Optimal Packing Characteristics of Rolled, Continuous Stationary‐Phase Columns

Abstract

Rolled, continuous stationary phases were constructed by tightly rolling and inserting a whole textile fabric into a chromatography column. This work reports the column performance, in terms of plate height, void fraction, and resolution, of 10 cellulose-based fabrics. The relation between fabric structural properties of yarn diameter, fabric count, fabric compressibility, and column performance are quantitated. General requirements, including reproducibility of packing, for choosing fabrics to make a good SEC column are identified. This research showed that the packed columns have an optimal mass of fabric that minimizes plate height and maximizes resolution, in a manner that is consistent with chromatography theory. Mass of material packed is then an important column parameter to consider when optimizing columns for the rapid desalting of proteins. Proteins were completely separated from salt and glucose in less than 8 min at a pressure drop less than 500 psi on the rolled, continuous stationary-phase columns. These results, together with stability and reproducibility, suggest potential industrial applications for cellulose-based rolled, continuous stationary-phase columns where speed is a key parameter in the production process.

Use of polymer fiber stationary phases for liquid chromatography separations: Part II – applications

“…Increases in packing density lead to broadening and peak asymmetry, presumably due to crimping of the interfiber channels. These observations are consistent with the findings of Ladisch and coworkers [24] who found that somewhat loosely packed, large yarns were preferred. The role of column diameter on peak characteristics demonstrates the directed flow patterns set up within the C-CP fiber column structure.…”

Roles of interstitial fraction and load conditions on the dynamic binding capacity of proteins on capillary‐channeled polymer fiber columns

“…This differs substantially from the case where fabrics consisting of yarns, made of multiple twisted filaments, have been packed in chromatographic columns. 21,23,28,29 As described by Ladisch and coworkers, 23 those structures provide a unique situation were different levels of porosity exist; intrafiber, between the fibers making up a given yarn (interfiber/intrayarn), and the interstices of the fabric structure (interyarn). Thus, different rates and modalities of protein mass transport exist at each level.…”

“…Perhaps the most substantive early works in this area were those of Ladisch and coworkers, who demonstrated the utility of textile fabrics to affect protein separation in a variety of chromatographic modes. 21,23,28,29 Those works, among other things, demonstrated the ability to affect high fluidic flow rates, without severe backpressure or solute mass transfer penalties. The key aspects here are a highly porous bed, with short inter-fiber diffusion distances, and a lack of intra-fiber porosity.…”

Rapid chromatography for evaluating adsorption characteristics of cellulase binding domain mimetics

“…The microstructure of individual cellulose fibrils that compose a cotton cellulose thread impart some size-exclusion capability to a rolled stationary-phase column, as can be seen in the near-baseline separation between Dextran (66.3 kDa) and sodium chloride (Li et al, 2002). However, cellulose binding, not size-exclusion effects, is the property of interest for mimetic binding domain candidates.…”

“…Cotton and cotton/polyester blend textiles in tightly rolled cylinders have been shown to make good continuous stationary-phase materials for rapid chromatography (Hamaker et al, 1996(Hamaker et al, , 1998Li et al, 2002). The material chosen for the adsorption chromatography screening was an untreated cotton print cloth, which is nearly 100% cellulose.…”