Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/26121
Type: Artigo de periódico
Title: Purification of recombinant aprotinin produced in transgenic corn seed: separation from CTI utilizing ion-exchange chromatography
Author: Azzoni, A. R.
Takahashi, K.
Woodard, S. L.
Miranda, E. A.
Nikolov, Z. L.
Abstract: Protein expression in transgenic plants is considered one of the most promising approaches for producing pharmaceutical proteins. As has happened with other recombinant protein production schemes, the downstream processing (dsp) of these proteins produced in plants is key to the technical and economic success of large-scale applications. Since dsp of proteins produced transgenically in plants has not been extensively studied, it is necessary to broaden the investigation in this field in order to more precisely evaluate the commercial feasibility of this route of expression. In this work, we studied the substitution of an IMAC chromatographic step, described in previous work (Azzoni et al., 2002), with ion-exchange chromatography on SP Sepharose Fast Flow resin as the second step in the purification of recombinant aprotinin from transgenic maize seed. The main goal of this second purification step is to separate the recombinant aprotinin from the native corn trypsin inhibitor. Analysis of the adsorption isotherms determined at 25°C under different conditions allowed selection of 0.020 M Tris pH 8.5 as the adsorption buffer. The cation-exchange chromatographic process produced a high-purity aprotinin that was more than ten times more concentrated than that generated using an IMAC step.
Subject: Recombinant aprotinin
Transgenic corn
Protease inhibitor purification
Downstream processing
Ion-exchange chromatography
Editor: Brazilian Society of Chemical Engineering
Rights: aberto
Identifier DOI: 10.1590/S0104-66322005000300001
Address: http://dx.doi.org/10.1590/S0104-66322005000300001
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0104-66322005000300001
Date Issue: 1-Sep-2005
Appears in Collections:Artigos e Materiais de Revistas Científicas - Unicamp

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