The primary production of an infectious recombinant Herpes Simplex Virus vaccine

• Published in: Biotechnology and bioengineering 1998-02, Vol.57 (3), p.262-271
• Main Authors: O'Keeffe, R., Johnston, M. D., Slater, N. K. H.
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Biotechnology; Cell culture; cell rupture; Cercopithecus aethiops; dextran sulphate; DISC HSV-2; Fundamental and applied biological sciences. Psychology; Glycerol; Health. Pharmaceutical industry; heparin; Herpes Simplex Virus type 2; Herpesvirus 2, Human - genetics; Herpesvirus 2, Human - immunology; Herpesvirus 2, Human - isolation & purification; Humans; Industrial applications and implications. Economical aspects; Microscopy, Electron; Monolayers; Osmosis; Production of active biomolecules; Proteins; Vaccines, Synthetic - biosynthesis; Vaccines, Synthetic - isolation & purification; Vaccins; Vero Cells; Viral Vaccines - biosynthesis; Viral Vaccines - isolation & purification; Virus Cultivation; virus release; Viruses
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/(SICI)1097-0290(19980205)57:3<262::AID-BIT2>3.0.CO;2-F

The production and extracellular release of a recombinant Herpes Simplex Virus (type 2) from monolayers of infected complementing Vero cells (CR2) are addressed. Growth and virus production conditions are identified that provide adequate virus titers with cell seeding densities and viral multiplicities of infection that could be reasonably handled in manufacturing. Harvesting by sonication of cell monolayers is shown to give the highest recovery of infectious virus (to 2.5 × 106 pfu/mL) but leads to process stream contamination by cellular proteins through the rupturing of cells (to 28 pg protein/pfu). By comparison, freeze‐thaw cycles and osmotic rupture by hypotonic saline or glycerol shock procedures yield only low virus recovery (typically