Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/195137
Type: Artigo de periódico
Title: Recombinant Aprotinin Produced In Transgenic Corn Seed: Extraction And Purification Studies.
Author: Azzoni, Adriano R
Kusnadi, Ann R
Miranda, Everson A
Nikolov, Zivko L
Abstract: Expression in transgenic plants is potentially one of the most economical systems for large-scale production of valuable peptide and protein products. However, the downstream processing of recombinant proteins produced in plants has not been extensively studied. In this work, we studied the extraction and purification of recombinant aprotinin, a protease inhibitor used as a therapeutic compound, produced in transgenic corn seed. Conditions for extraction from transgenic corn meal that maximize aprotinin concentration and its fraction of the total soluble protein in the extract were found: pH 3.0 and 200 mM NaCl. Aprotinin, together with a native corn trypsin inhibitor (CTI), was captured using a tryspin-agarose column. These two inhibitors were separated using an agarose-IDA-Cu2+ column that proved to efficiently absorb the CTI while the recombinant aprotinin was collected in the flowthrough with purity of at least 79%. The high purity of the recombinant aprotinin was verified by SDS-PAGE and N-terminal sequencing. The overall recombinant aprotinin recovery yield and purification factor were 49% and 280, respectively. Because CTI was also purified, the recovery and purification process studied has the advantage of possible CTI co-production. Finally, the work presented here introduces additional information on the recovery and purification of recombinant proteins produced in plants and corroborates with past research on the potential use of plants as biorreactors.
Subject: Adsorption
Aprotinin
Buffers
Chromatography, Affinity
Copper
Gene Expression Regulation, Plant
Hydrogen-ion Concentration
Plant Proteins
Plants, Genetically Modified
Recombinant Proteins
Seeds
Sensitivity And Specificity
Sodium Chloride
Temperature
Zea Mays
Rights: fechado
Identifier DOI: 10.1002/bit.10408
Address: http://www.ncbi.nlm.nih.gov/pubmed/12226858
Date Issue: 2002
Appears in Collections:Unicamp - Artigos e Outros Documentos

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