DEVELOPMENT OF SUPERMACROPOROUS MATRICES WITH ETHYLAMINE IMMOBILIZED FOR PURIFICATION OF PROTEINS

The demand of food industries for active bio composites is increasing. Because of this, the development of monolithic polymeric structures is promising, with cryogels being the most prominent. These adsorbents require modifications, which aim to increase their purification capacity. An alternative is the hydrophobic interaction technique, which is based on the insertion of ligands that interact specifically with the nonpolar portions of the proteins, generating compounds with a high degree of purity. Therefore, the aim of this study is to develop a supermocroporous cryogel activated with ethylamine, aimed at purifying proteins.
The cryogels were synthesized from an aqueous solution containing Acrylamide, N, N'-methylenebisacrylamide and allyl glycidyl, totaling 7% of monomers. Added ammonium persulfate and N, N, N ', N'-tetramethylethylenediamine. The solution was poured into 5 ml plastic syringes and immersed in a thermostatic bath at a temperature of 12 ° C for 24 h. Afterwards, they were dried in an oven at 60ºC. They were then functionalized with ethylamine and glutaraldehyde. For the evaluation of efficiency, the cryogels were placed in contact with a solution of Bovine Serum Albumine at 1 mg.mL-1 prepared in sodium phosphate buffer containing different concentrations of sodium sulfate (1 Molar and 1.5 Molar).The adsorbed amount was determined by the Bradford method.
The adsorbent produced had a higher adsorption capacity (31.76 ± 0.03 mg of protein. g-1 of cryogel) when placed in contact with the solution containing the highest concentration of salt (1.5 Molar). The use of high concentrations of salt promotes the interactions between ligand-protein, and consequently increases the retention of proteins. Therefore, the results show that the salt molecules increase the surface tension of the solution, stimulating the exposure of the internal hydrophobic regions present in the protein. Thus, the cryogels produced have potential for use in protein purification processes through hydrophobic interaction chromatography.