Artículos de revistas
Negative chromatography on agarose-TREN as a technique for purification of protein spiked in soybean seeds extract
Registro en:
Process Biochemistry. Elsevier Sci Ltd, v. 47, n. 12, n. 2255, n. 2261, 2012.
1359-5113
WOS:000313851700073
10.1016/j.procbio.2012.08.023
Autor
Bresolin, IRAP
Bresolin, ITL
Miranda, EA
Bueno, SMA
Institución
Resumen
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Alkyl amines and polyamines have been used as ligands for protein purification by mixed-mode chromatography. The adsorption of proteins onto these ligands seems to be governed by multiple effects such as electrostatic, hydrophobic, and affinity interactions. In this work we investigated the adsorption of proteins extracted from soybean onto the adsorbent agarose-Tris(2-aminoethyl)amine (TREN). The effects of flow rate, buffer system, and extract concentration on the capture of proteins extracted from soybean were evaluated. Experiments using Mes at pH 6.5 as adsorption buffer allowed the adsorption of almost the totality of native soybean protein with a dynamic adsorption capacity of 13.50 mg mL(-1) adsorbent. Experiments with human IgG (pl in the range of 5.8-9.0) and human serum albumin (HSA, pl of 4.9) spiked into these extracts lead to the conclusion that electrostatic forces play a major role in the interaction between protein and agarose-TREN. Based on this work, negative chromatography with agarose-TREN should be considered as a method for purification of basic recombinant protein produced in transgenic soybean seeds. (C) 2012 Elsevier Ltd. All rights reserved. 47 12 2255 2261 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)