Purification of recombinant aprotinin produced in transgenic corn seed: separation from CTI utilizing ion-exchange chromatography
| Autor: | S. L. Woodard, K. Takahashi, Everson Alves Miranda, Adriano R. Azzoni, Z. L. Nikolov |
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| Jazyk: | angličtina |
| Rok vydání: | 2005 |
| Předmět: |
Tris
Genetically modified maize Chromatography Downstream processing Chemistry General Chemical Engineering Ion chromatography Ion-exchange chromatography lcsh:TP155-156 Genetically modified crops Protease inhibitor purification law.invention Recombinant aprotinin chemistry.chemical_compound Adsorption law Recombinant DNA medicine Aprotinin Transgenic corn lcsh:Chemical engineering medicine.drug |
| Zdroj: | Brazilian Journal of Chemical Engineering, Vol 22, Iss 3, Pp 323-330 (2005) Brazilian Journal of Chemical Engineering, Volume: 22, Issue: 3, Pages: 323-330, Published: SEP 2005 Brazilian Journal of Chemical Engineering v.22 n.3 2005 Brazilian Journal of Chemical Engineering Associação Brasileira de Engenharia Química (ABEQ) instacron:ABEQ |
| ISSN: | 1678-4383 0104-6632 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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