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Hum Vaccin Immunother. 2013 Oct;9(10):2211-5. doi: 10.4161/hv.25048. Epub 2013 Jul 30.

Antibiotic-free production of a herpes simplex virus 2 DNA vaccine in a high yield cGMP process.

Author information

1
VGXI, Inc.; The Woodlands, TX USA.
2
Coridon Pty Ltd; Level 3, Translational Research Institute; Woolloongabba, QLD Australia.
3
Nature Technology Corporation; Lincoln, NE USA.

Abstract

Two DNA vaccine plasmids encoding Herpes simplex virus type 2 (HSV-2) glycoprotein D, NTC8485-O2-gD2 and NTC8485-O2-UgD2tr, were produced at large scale under current good manufacturing practice (cGMP) for use in a Phase I human clinical trial. These DNA vaccines incorporate the regulatory agency compliant, minimal, antibiotic-free (AF) NTC8485 mammalian expression vector. Plasmid yields of>1 g/L were achieved using the HyperGRO™ fed-batch fermentation process, with successful scale up from 10 L process development scale to 320 L culture volume for cGMP production. The DNA vaccines were purified using a low residence time, high shear lysis process and AIRMIX(TM) technology, followed by chromatographic purification. This combination of optimized plasmid vector, high yield upstream production, and efficient downstream purification resulted in purified HSV-2 DNA vaccines with>99% total supercoiled plasmid, ≤ 0.2% RNA, ≤ 0.1% host cell genomic DNA, and ≤ 0.1 endotoxin units per mg.

KEYWORDS:

DNA vaccine; Escherichia coli; HSV-2; antibiotic-free; downstream processing; fermentation; plasmid

PMID:
23899469
PMCID:
PMC3906406
DOI:
10.4161/hv.25048
[Indexed for MEDLINE]
Free PMC Article
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