Rolled stationary phases: Dimensionally structured textile adsorbents for rapid liquid chromatography of proteins

A woven textile fabric, consisting of 60% cotton/40% polyester, tightly rol led in a cylindrical configuration, has a three-dimensional structure with sufficient hydrodynamic stability to withstand interstitial eluent velociti es of up to 300 cm/min when packed into standard liquid chromatography colu mn assemblies. Demonstration of the pressure stability of the cotton/ polye ster fabric was followed up with experiments in which the cotton (cellulose ) portion was derivatized and the fabric evaluated for chromatography of pr oteins. When derivatized to give a (diethylamino)ethyl (DEAE) anion exchang er, a velocity independent plate height of 2 mm, a static capacity of 115 m g of bovine serum albumin/g of stationary phase, and a dynamic protein load ing capacity which decreases only 25% over an 800% increase in mobile-phase velocity from 6.7 to 54 cm/min was achieved. The fibers that make up the s tationary phase have a relatively nonporous structure which minimizes pore diffusional effects. A protein separation of Cytochrome C from beta-lactogl obulin A is shown to be completed by ion-exchange chromatography in less th an 10 min using an NaCl step gradient. Gradient chromatography of a hen egg white shows resolution of the proteins into two major components (lysozyme and ovalbumin) as well as two minor ones. A size exclusion separation of P EG 20 000 from glucose requires only 90 s. These characteristics, together with the ability of the cellulose-based stationary phase to withstand rapid flow rates, indicate that this type of stationary phase has potential for applications where chromatography using DEAE-cellulose particles has proven successful.