Document Detail


Purification of an acidic recombinant protein from transgenic tobacco.
MedLine Citation:
PMID:  17787005     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Tobacco has proven to be a promising alternative for the production of recombinant therapeutic proteins and offers numerous advantages over other plants as a host system. However, the recovery and purification steps needed to obtain a protein at high recovery and purity have not been well investigated. In this study, a process was developed to purify a model acidic protein, recombinant beta-glucuronidase (rGUS) from transgenic tobacco leaf tissue, in three main steps after extraction: polyelectrolyte precipitation, hydrophobic interaction chromatography (HIC), and hydroxyapatite chromatography (HAC). Using this three-step process, up to 40% of the initial rGUS activity could be recovered to near homogeneity as judged by SDS-PAGE. This work demonstrates that acidic recombinant proteins expressed in tobacco may be purified to high yield with high purity in a minimal amount of steps that are suitable for scale-up. Furthermore, the general steps used in this process may suggest that a wide variety of acidic recombinant proteins may be purified in a similar manner from transgenic tobacco or other leafy crops.
Authors:
Chris Holler; Chenming Zhang
Related Documents :
10419825 - Expression and purification of recombinant human zona pellucida proteins.
20545445 - Recombinant n-terminal part of bovine herpesvirus 1 icp27 protein: its preparation, pur...
12215815 - Isolation of a novel thermus thermophilus metal efflux protein that improves escherichi...
9882565 - Defective vaccinia virus as a biologically safe tool for the overproduction of recombin...
14757845 - Molecular cloning and characterization of the von hippel-lindau-like protein.
2235995 - Distinct character in hydrophobicity of amino acid compositions of mitochondrial proteins.
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  99     ISSN:  1097-0290     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2008 Mar 
Date Detail:
Created Date:  2008-01-31     Completed Date:  2008-02-21     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  902-9     Citation Subset:  IM    
Copyright Information:
Copyright 2007 Wiley Periodicals, Inc.
Affiliation:
Department of Biological Systems Engineering, Virginia Polytechnic Institute and State University, 210 Seitz Hall, Blacksburg, Virginia 24061, USA.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Descriptor/Qualifier:
Chromatography / methods
Glucuronidase / isolation & purification*,  physiology*
Plant Leaves / metabolism*
Plants, Genetically Modified / metabolism*
Recombinant Proteins / isolation & purification*,  metabolism*
Tobacco / physiology*
Chemical
Reg. No./Substance:
0/Recombinant Proteins; EC 3.2.1.31/Glucuronidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


Previous Document:  Ex vivo and in vivo evaluation of (2R,3R)-5-[(18)F]-fluoroethoxy- and fluoropropoxy-benzovesamicol, ...
Next Document:  Electrooptical measurements for monitoring metabolite fluxes in acetone-butanol-ethanol fermentation...