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  4. Overcoming inefficient secretion of recombinant VEGF-C in baculovirus expression vector system by simple purification of the protein from cell lysate
 
research article

Overcoming inefficient secretion of recombinant VEGF-C in baculovirus expression vector system by simple purification of the protein from cell lysate

Klaus, Tomasz
•
Kulesza, Malgorzata
•
Bzowska, Monika
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2015
Protein Expression And Purification

The first reports about successfully expressed recombinant proteins with the use of a baculovirus vector were published over 30 years ago. Despite the long time of refining this expression system, early problems with the production of baculovirus-derived secretory proteins are still not satisfactorily solved. The high expression level driven by baculoviral promoters often does not result in the desired yield of secreted recombinant proteins, which frequently accumulate inside insect cells and are only partially processed. During our attempts to produce vascular endothelial growth factor C (VEGF-C) with the use of a baculovirus vector we also faced an inefficient secretion of the recombinant protein to culture medium. We were not able to improve the outcome and obtain an acceptable concentration of VEGF-C in the medium by changing the culture conditions or utilizing different signal peptides. However, as a significant amount of native VEGF-C was detected inside the baculovirus-infected cells, we developed a simple method to purify recombinant, glycosylated VEGF-C from a lysate of the cells. The presented results indicate that the lack of a secretory protein in the insect cell culture medium after baculovirus infection does not necessarily signify failure in the production of the protein. As demonstrated by us and contrary to generally accepted views, the lysate of baculovirus-infected cells may constitute a valuable source of the biologically active, secretory protein. (C) 2015 Elsevier Inc. All rights reserved.

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Type
research article
DOI
10.1016/j.pep.2015.03.001
Web of Science ID

WOS:000353313500022

Author(s)
Klaus, Tomasz
Kulesza, Malgorzata
Bzowska, Monika
Wyroba, Barbara
Kilarski, Witold W.
Bereta, Joanna
Date Issued

2015

Publisher

Academic Press Inc Elsevier Science

Published in
Protein Expression And Purification
Volume

110

Start page

151

End page

158

Subjects

Baculovirus expression vector system

•

Secretion

•

Vascular endothelial growth factor C

•

VEGF-C

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
IBI-SV  
Available on Infoscience
May 29, 2015
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/114158
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