title: Antibiotic-free production of a herpes simplex virus 2 DNA vaccine in a high yield cGMP process.
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.
authors.affiliation: VGXI, Inc.; The Woodlands, TX USA.
authors.forename: Jared
authors.initials: J
authors.lastname: Nelson
authors.affiliation: VGXI, Inc.; The Woodlands, TX USA.
authors.forename: Stephen
authors.initials: S
authors.lastname: Rodriguez
authors.affiliation: Coridon Pty Ltd; Level 3, Translational Research Institute; Woolloongabba, QLD Australia.
authors.forename: Neil
authors.initials: N
authors.lastname: Finlayson
authors.affiliation: Nature Technology Corporation; Lincoln, NE USA.
authors.forename: Jim
authors.initials: J
authors.lastname: Williams
authors.affiliation: Nature Technology Corporation; Lincoln, NE USA.
authors.forename: Aaron
authors.initials: A
authors.lastname: Carnes
mesh_terms: D000900:Anti-Bacterial Agents; D005822:Genetic Vectors; D022301:Herpes Simplex Virus Vaccines / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D018258:Herpesvirus 2, Human / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D012641:Selection, Genetic; D013678:Technology, Pharmaceutical / Q000379:methods*; D019444:Vaccines, DNA / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
chemical_list: D000900:Anti-Bacterial Agents; D022301:Herpes Simplex Virus Vaccines; D019444:Vaccines, DNA
keywords: DNA vaccine; Escherichia coli; HSV-2; antibiotic-free; downstream processing; fermentation; plasmid
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