By Ralf Pörtner
Animal cellphone Biotechnology: equipment and Protocols, moment variation constitutes a entire handbook of state of the art and new ideas for developing mammalian phone strains for creation of biopharmaceuticals, and for optimizing serious parameters for phone tradition contemplating the complete cascade from lab to ultimate creation. the quantity is split into 5 elements that mirror the methods required for various phases of construction. partially I, simple recommendations for institution of construction phone traces are addressed, in particular transduction strategies, cells for gene treatment and antibody creation. half II addresses uncomplicated cultivation strategies, resembling microcarrier tradition and encapsulation.
Part III covers mobile characterization and research, together with move cytometric purposes, NMR-based concepts, and biochemical and cytometric recommendations. half IV info cultivation concepts, similar to disposable bioreactors, hole fiber cellphone tradition, mounted mattress reactors, and configuration of bioreactors. half V covers downstream recommendations reminiscent of membrane filtration suggestions, whereas half VI describes exact purposes, together with retroviral vectors.
Animal mobilephone Biotechnology: tools and Protocols, moment variation presents a compendium of strategies for scientists in commercial and study laboratories that use mammalian cells for biotechnology purposes.
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Additional resources for Animal Cell Biotechnology: Methods and Protocols (Methods in Biotechnology)
Miller, D. , Garcia, J. , and Lynch, C. M. (1993) Use of retroviral vectors for gene transfer and expression. Methods Enzymol. 217, 581–599. 15. , et al. (1996) In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 272, 263–267. 20 Fluri and Fussenegger 16. Bukovsky, A. , Song, J. , and Naldini, L. (1999) Interaction of human immunodeficiency virus-derived vectors with wild-type virus in transduced cells. J. Virol. 73, 7087–7089. 17. Danthinne, X. and Imperiale, M.
Typical titers are in the range of 1–6 × 1010 virus particles (VPs) per mL. A recent study using the HeLaS3 human tumor cell line for propagating recombinant adenoviruses demonstrated yields of 6 × 1011 VP/mL and showed that this productivity could be maintained at pilot (70 L) scales (58). Successful large-scale production for all cell lines is dependent on optimization of the culture conditions and the best mode of operation. For more details, see Kamen and Henry (56) and Warnock et al. (59).
However, the maximum specific virus titer remains inferior to T-flask cultures (54). Hydrodynamic shear forces are believed to affect cellular receptor levels and thus hinder virus entry into HEK293 cells at high multiplicity of infection (MOI). Additionally, HEK293 cells have been cited as having a low ability to colonize microcarriers because of poor attachment (40). Suspension cultures are more appealing for large-scale production because of the ease of operation and scale-up. C6® cells can be grown in stirred tank bioreactors, and, depending on the mode of operation, cell densities are in excess of 5 × 106 cells/mL at scales from 3 to 20 L (42,56,57).
Animal Cell Biotechnology: Methods and Protocols (Methods in Biotechnology) by Ralf Pörtner