Negative chromatography on agarose-TREN as a technique for purification of protein spiked in soybean seeds extract

Negative chromatography on agarose-TREN as a technique for purification of protein spiked in soybean seeds extract

Author Antunes Pereira Bresolin, Iara Rocha Google Scholar
Lazzarotto Bresolin, Igor Tadeu Autor UNIFESP Google Scholar
Miranda, Everson Alves Google Scholar
Alves Bueno, Sonia Maria Google Scholar
Institution Universidade Estadual de Campinas (UNICAMP)
Universidade Federal de São Paulo (UNIFESP)
Abstract 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 B.V. All rights reserved.
Keywords Tris(2-aminoethyl)amine (TREN)
Mixed-mode chromatography
Negative chromatography
Soybean extract
Downstream processing
Language English
Sponsor Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Date 2012-12-01
Published in Process Biochemistry. Oxford: Elsevier B.V., v. 47, n. 12, p. 2255-2261, 2012.
ISSN 1359-5113 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 2255-2261
Origin http://dx.doi.org/10.1016/j.procbio.2012.08.023
Access rights Closed access
Type Article
Web of Science ID WOS:000313851700073
URI http://repositorio.unifesp.br/handle/11600/35573

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