Abstract
The baculovirus-insect cell expression system is an important technology for the production of recombinant proteins and baculovirus-based biopesticides. Budded virus titration is critical when scaling up baculovirus production processes in suspension cultures, to ensure reproducible infections, especially when a low multiplicity of infection (MOI) is applied. In this study, a simple suspension culture titration (SCT) assay was developed that involves accurate measurements of the initial cell densities (ICDs) and peak cell densities (PCDs) of an infected culture, from which the MOI and hence the virus inoculum infectious titer can be estimated, using the established Power-Nielsen baculovirus infection model. The SCT assay was assessed in parallel with two adherent culture-based assays (MTT and AlamarBlue) for the Heliothine baculovirus HaSNPV, and was shown to be more objective, time-efficient and reproducible. The model predicted a linear correlation between log(PCD/ICD) and log(MOI), hence an alternative model-independent SCT assay was also developed, which relies on a well-replicated standard curve relating suspension culture-derived PCD/ICD ratios with plaque or endpoint assay-derived MOIs. Standard curves with excellent lin...Continue Reading
References
Jan 1, 1992·Cytotechnology·L K NielsenP F Greenfield
Nov 1, 1994·Biotechnology Progress·J ZhangK Iatrou
Jan 1, 1997·Biotechnology Progress·K U Dee, M L Shuler
Apr 1, 1999·Biotechnology and Bioengineering·L C ChanS Reid
May 1, 1999·Journal of Invertebrate Pathology·S Chakraborty, S Reid
Oct 12, 2000·Journal of Invertebrate Pathology·D E Lynn
Jan 12, 2002·Biotechnology and Bioengineering·Matthew RosinskiLars Keld Nielsen
Sep 25, 2002·Journal of Virological Methods·Chun Fang ShenAmine Kamen
Mar 12, 2004·Annual Review of Entomology·F Moscardi
May 7, 2005·Nature Biotechnology·Thomas A KostDonald L Jarvis
Jun 11, 2005·Biotechnology and Bioengineering·Richard Haas, Lars K Nielsen
Jun 28, 2005·Journal of Theoretical Biology·Giora EndenJosé C Merchuk
Nov 1, 2005·Biotechnology Advances·Boguslaw SzewczykMarlinda Lobo de Souza
Feb 4, 2006·Biotechnology Progress·Hasnaa JorioAmine Kamen
May 24, 2006·Journal of Virological Methods·Márcia R S PedriniLeslie C L Chan
Feb 10, 2007·Journal of Biotechnology·António RoldãoR Oliveira
Apr 5, 2007·Nature Protocols·Vasantharajan JanakiramanSomasekar Seshagiri
Aug 5, 1992·Biotechnology and Bioengineering·C D De GooijerJ Tramper
Sep 5, 1994·Biotechnology and Bioengineering·J F PowerL K Nielsen
Jun 5, 1997·Biotechnology and Bioengineering·K U Dee, M L Shuler
May 5, 2009·Virology·Tilo SengerMartin Müller
May 16, 2009·Journal of Virological Methods·António RoldãoPaula M Alves
Jan 27, 2010·The Journal of Gene Medicine·Jimmy A MenaAmine A Kamen
Oct 26, 2010·Journal of Virological Methods·Matthew M FerrisKathy L Rowlen
Feb 1, 2011·Vaccine·R BaxterM M Cox
Jun 7, 2011·Biotechnology Progress·Márcia R S PedriniLeslie C L Chan
Aug 3, 2011·Expert Review of Vaccines·Jimmy A Mena, Amine A Kamen
Jan 1, 1996·Cytotechnology·J F Power, L K Nielsen
Citations
Jul 26, 2013·Archives of Virology·Leila MatindoostSteven Reid
May 8, 2015·Viruses·Leila MatindoostSteve Reid
Jun 21, 2015·Archives of Virology·Hoai T HuynhSteven Reid
Sep 11, 2016·Applied Microbiology and Biotechnology·Quan NguyenSteven Reid
Oct 22, 2016·Biotechnology Progress·Ignacio EberhardtJuan Daniel Claus