Document Detail

Characterization of green-tissue protein extract from alfalfa (Medicago sativa) exploiting a 3-D technique.
MedLine Citation:
PMID:  19746403     Owner:  NLM     Status:  MEDLINE    
There is a growing interest of pharmaceutical companies for plant-based production systems. To facilitate the general acceptance of plants as bioreactors, the establishment of efficient downstream operations is critical. It has been proposed that a better understanding of the properties of the contaminant proteins can benefit downstream processing design and operation. The coupled application of 2-DE with aqueous two-phase partitioning has been suggested as a practical 3-D method to characterize potential contaminant proteins from plant extracts. The application of this novel 3-D approach to a complex protein extract from alfalfa (Medicago sativa) containing a model recombinant protein (human granulocyte colony stimulating factor (hG-CSF)) resulted in the quantification of 55 protein spots. The 3-D properties (M(r), pI, and K(p)) obtained for 17 proteins comprising 69% of the alfalfa proteins, allowed the proposal of a prefractionation step as well as the identification of the target molecule (rG-CSF) from bulk of alfalfa proteins. The information obtained from this experimental approach was useful for the identification of the potential contaminant proteins that will occur in alfalfa when this plant is used as a host for recombinant proteins. Additionally, this method will assist in the design of adequate purification strategies for recombinant proteins expressed in alfalfa green tissue.
Oscar Aguilar; Charles E Glatz; Marco Rito-Palomares
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of separation science     Volume:  32     ISSN:  1615-9314     ISO Abbreviation:  J Sep Sci     Publication Date:  2009 Sep 
Date Detail:
Created Date:  2009-09-15     Completed Date:  2010-01-26     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101088554     Medline TA:  J Sep Sci     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  3223-31     Citation Subset:  IM    
Departamento de Biotecnolog??a e Ingenier??a de Alimentos, Centro de Biotecnolog??a, Tecnol??gico de Monterrey, Campus Monterrey, Monterrey, NL, M??xico.
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MeSH Terms
Chemical Fractionation
Electrophoresis, Gel, Two-Dimensional
Medicago sativa / chemistry*
Plant Extracts / isolation & purification*
Plant Proteins / isolation & purification*
Reg. No./Substance:
0/Plant Extracts; 0/Plant Proteins

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