COMPOSITE CELLULOSE FIBRE FOR AFFINITY CHROMATOGRAPHY APPLICATION

Affinity chromatographic supports are nowadays one of the most frequently used and expensive consumable materials for protein purification at the laboratory and industrial scales. The introduction of cost-effective materials is an important issue to address in order 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 purpose. Nevertheless, fibre insolubility and low chemical reactivity are the major drawbacks preventing the use of this material for protein chromatography. In this work, a composite prepared from polymethacrylate/cellulose fibres was used for the preparation of chromatographic materials containing immobilised proteins as adsorptive ligands. A green fluorescent protein and a protein A-similar ligand, containing cysteine in the terminal tags, were immobilised onto the fibres by epoxy-thiol chemistry. Buffer salt, pH, reaction time and pre-swelling procedure were optimised. The materials were characterised by fluorescence and electron microscopy, in addition to the binding capacity of the adsorbent materials.