Novel Porous Monolithic Column Using Poly(high internal phase emulsion) Methacrylate as Materials for Immunoglobulin Separation Performance on HPLC

Abstract

AbstractA novel porous polymeric monolithic column based on poly(high internal phase emulsion) methacrylate monolith was prepared and applied to fast separation of proteins. The block copolymer chemistry of high internal phase emulsions was used in the experiment. These unique properties, together with high porosity, good mechanical property, chemical modification of the surface make themselves superior in monolithic medium applications. Morphology of the monolithic material was studied by scanning electron microscopy. The stability of the emulsion and the load of hydroxyl groups–the active group of the monolithic column were investigated. Additionally, the capabilities of separation of this column in conjunction with high performance liquid chromatography (HPLC) were investigated. Immunoglobulin was separated from human plasma and chicken egg yolk with high resolution on the hydrophobic interaction chromatographic column in a short time. The effects of pH and concentration of mobile phase (buffer) on the elution of immunoglobulin were investigated. Moreover, fast separation of a two mode protein mixture (α‐amylase and lysozyme) on the monolith was achieved within 1.5 min at a velocity of 1445 cm·h−1. As a result, good separation was achieved, and stable low back pressure drop was ensured at high throughput elution with an even longer column.