Rolled, continuous stationary phase columns from textile fabrics: Optical packing and protein separation.

摘要

Rolled, continuous stationary phases were constructed by tightly rolling and packing a whole piece of textile woven fabric into a chromatography column. Columns packed in this manner possess important features such as low intraparticle mass transfer and high pressure endurance. These properties, as reflected by a reduction of residence time within a column, show promise in decreasing protein purification costs for biotechnology and pharmaceutical companies. This technique, once commercialized, will create a new value-added market for textile fabrics. The purpose of this thesis is to demonstrate the capabilities of rolled, continuous stationary phase columns in size exclusion chromatography (SEC) separation, ion exchange chromatography (IEC) separation, and pore characterization of cellulase enzyme treated cotton.;SEC stationary phase columns were prepared from ten cotton-based fabrics with different yarn diameter, fabric count, fabric compressibility, fabric weave, and/or different chemical treatment(s). The optimal packing parameter with respect to column mass was identified to be 7.5--8 grams for a column with the dimension 180 mm by 10 mm, length by inner diameter. Evaluation of resolution, plate height, and void fraction of the packed column revealed the general requirements of a good SEC separation for a fabric to be large yarns, a small fabric count, and a plain weave with napping. A complete separation of protein from salt was achieved on a series of cotton flannel stationary phases that were 510 mm long and 10 mm in diameter. Cotton flannel anion exchangers with a static protein capacity of 45 mg/g were prepared from a quaternary ammonium 3-chloro-2-hydroxypropyl trimethylammonium chloride (CHPTAC) with a repeated padding method. Columns packed with CHPTAC derivatized cotton flannel completely resolved BSA and lysozyme. A complete separation of BSA from lysozyme on a scoured cotton flannel column suggested that at pH 8.0, the fabric is negatively charged on the surface. With the rolled, continuous stationary phase technique, pore volume and surface area of cellulase enzyme treated cotton were characterized. No increase in pore volume or surface area was observed after enzyme hydrolysis, which agrees with the non-increasing dye uptake of enzyme treated fabric.