Composite cellulose fibre for affinity chromatography application

Abstract

Affinity chromatographic supports is nowadays one of the most used and expensive consumable material for protein purification at the laboratory and industrial scale. The introduction of cost-effective materials is an important issue to address to spread the usefulness of this technology. Cotton fibres are a highly available natural material with excellent mechanical and structural properties which can be used for this propose. Nevertheless, fibre insolubility and low chemical reactivity are the major drawbacks to use this material for protein chromatography.In this work, a composite material prepared from polymethacrylate/cellulose fibres were used for the preparation of chromatographic materials containing immobilised proteins as adsorptive ligands. A green fluorescent protein and a protein A simil ligand containing cysteine in the terminal tags were immobilised onto the fibres by the epoxy-thiol chemistry. Buffer salt, pH, reaction time and pre-swelling procedure were optimised. Materials were characterised by fluorescence and electron microscopes, in addition to the binding capacity of the adsorbent materials.