Abstract
Disabled Infectious Single Cycle (DISC) HSV-2 is cultured in a complimentary cell line (CR2) in microcarrier cultures up to a 15 L scale. Virus is released from the cells by the addition of a hypotonic saline solution and using low-pressure disruption technique. The resultant material must be further purified to decrease contaminating host cell protein and DNA.
The aim of our study was to demonstrate that incubation with heparin at the point of harvest could beused to release the virus from the cells, providing a favourable alternative to the cell disruption method.
Infected cultures were harvested at approxmately 100%CPE by incubation with porcine mucosal heparin.
Virus titres equivalent to those obtained by disruption techniques were consistently obtained using heparin incubation. The intact cells were easily separated from the released virus in the supernatant. The levels of contaminants were reduced for the start of further purification steps.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Corey L. (1993). Herpes Simplex virus infections duiring the decade since licensureof acyclovir J. bled. Virol.. Suppl.: 1:7–12.
McLean CS, Erturk M. Jennings R, Ni Challanain D. Minson AC. Boursnell MEG und Inglis SC (1994). Protective vaccination against primary and recurrent disease caused by herpes simplex virus type 2 using a genetically disabled by herpes simplex virus type I virus J. Inf. Dis.: 170. 1100–1109.
Boursnell MEC, Entwistle C. Blakely D. Roberts C. Duncan IA, Chisholm SE, Martin GM. Jennings R, Ni Challanain D, Sobek I and McLean CS. (1997). A genetically inactivated Herpes Simplex virus type 2 2 HSV) vaccine provides effective protection against primary and recurrent HSV desease J. Inf Dis.: 175: 16–25.
Zecchini TA and Smith RJ. (1997). Production of high titre DISC-HSV-2 from a microcarrier culture, In New Developments and New Applications in Cell Technology Preceedings of the 15th ESACT meeting Eds. Merton O-W, Perrin P and Grifffiths B. Kluwer Academic Publishers
Degouys V, Montoisy C. Wanderpepen M-P, Togasaki RK, Surzcki SJ. Segault V and Miller AOA. (1197). “Bionebulisation” an alternative to Dounce homogenisation for the preparation of mammalian cells nuclei in bulk. pp271–275 in Animal Cell Technology (From Vaccines to Genetic Medicine) Eds. Carronda M.J.T.. Griffiths B. and Moreira L.P.: Kluwer Academic Publishers Dordrecht Holland.
O’Keeffe R. Johnston MD, Slater NKH. (1998) The primary preparation of an infectious desease Herpes Simplex virus vaccine Biotech. Bioeng.: 57: No. 3: 262–271.
Author information
Authors and Affiliations
Editor information
Rights and permissions
Copyright information
© 2002 Kluwer Academic Publishers
About this chapter
Cite this chapter
Wright, P.A., Zecchini, T.A., Smith, R.J., O’Keefe, R.S., Slater, N.K. (2002). Incubation with Heparin as an Improved Harvest Method for High Titre Disabled Infectious Single Cycle (DISC) HSV-2 from Microcarrier Cultures. In: Ikura, K., Nagao, M., Masuda, S., Sasaki, R. (eds) Animal Cell Technology: Challenges for the 21st Century. Springer, Dordrecht. https://doi.org/10.1007/0-306-46869-7_20
Download citation
DOI: https://doi.org/10.1007/0-306-46869-7_20
Publisher Name: Springer, Dordrecht
Print ISBN: 978-0-7923-5805-3
Online ISBN: 978-0-306-46869-8
eBook Packages: Springer Book Archive