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.

Keywords: :

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

Disclosure of Potential Conflicts of Interest

Jared Nelson and Stephen Rodriguez have a financial interest in VGXI, Inc. Neil Finlayson has a financial interest in Coridon Pty Ltd. Aaron E Carnes and James A Williams have a financial interest in Nature Technology Corporation.