fulltext.study @t Gmail

Improving baculovirus production at high cell density through manipulation of energy metabolism

Paper ID Volume ID Publish Year Pages File Format Full-Text
31759 44836 2010 14 PDF Available
Title
Improving baculovirus production at high cell density through manipulation of energy metabolism
Abstract

The insect cells/baculovirus system is well recognized as a safe and suitable technology to produce heterologous proteins, vaccines and vectors for gene therapy. Efficient and robust production processes, able to deliver higher product concentrations, are however still needed to cope with increased requirements for large-scale manufacture. The work herein presented describes a combined experimental and modelling effort to quantify and environmentally manipulate the metabolism of Spodoptera frugiperda cells, targeting high cell density production of baculovirus vectors with potential application in human gene therapy. Culture medium supplementation with pyruvate or α-ketoglutarate at the time of infection resulted in 6–7-fold higher specific baculovirus yields at high cell density when compared to control cultures. This pushed volumetric titers to levels higher than classical low cell density infections. A quantitative description of intracellular pathways is provided using metabolic flux analysis; a direct stimulation of carbon flow through the tricarboxylic acids cycle was observed. Analysis of flux partitioning coefficients at the pyruvate and α-ketoglutarate branch-points further revealed a metabolic transition to a more energetically active state, which was confirmed by increased intracellular adenosine triphosphate generation rates. These results represent a cost-efficient and scalable strategy for high cell density production of recombinant baculovirus vectors.

Keywords
Sf-9 insect cells; High cell density production; Metabolic flux analysis; Pyruvate; α-Ketoglutarate; Energy metabolism; Baculovirus; Gene therapyAcoA, acetyl-coenzyme A; ADP, adenosine diphosphate; AlaAT, alanine aminotransferase; Amm, ammonia; Arg, argin
First Page Preview
Improving baculovirus production at high cell density through manipulation of energy metabolism
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Metabolic Engineering - Volume 12, Issue 1, January 2010, Pages 39–52
Authors
, , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us