Protein chromatography using a continuous stationary phase

A continuous stationary phase consisting of yarns woven into a fabric is rolled and packed into mechanically stable liquid chromatography columns. This work utilized yarns having a characteristic width of 200-400 μm, made from 10-20-μm fibers consisting of 95% poly(m-phenylene isophthalamide) and 5% poly(p-phenylene terephthalamide). Although loadings on this stationary phase were low at 4 mg/g for bovine serum albumin and 6 mg/g for β-galactosidase, this material shows the interesting characteristic of a leveling off of plate height at mobile phase velocities of 30-80 cm/min. This phenomenon is explained on the basis of a coupling argument whereby a fraction of the mobile phase flows through the intramatrix pore space, and convective transport through the pore space dominates transport by diffusion. A modified Van Deemter expression is derived and shown to fit plate height data for polyethylene glycol standards having molecular weights of 200 and 20 000. The characteristics of this continuous stationary phase at high eluent velocities are discussed conditions which give separation of immunoglobulin G, bovine serum albumin, insulin and β-galactosidase in 12 min are described.