Abstract
An on-line fluorescence sensor prototype was constructed to monitor the production of the green fluorescent protein (GFP) by 293S cells infected with a recombinant adenovirus vector (rAdV) containing the GFP gene. Fluorescence was correlated to GFP concentration and therefore to viral protein expression, but not to rAdV production. The sensor signal can also be used to compute the GFP production rate, which predicts well the occurrence of maximum viral titer.
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References
Albano C, Randers-Eichhorn L, Bentley W, Rao G (1998) Green fluorescent protein as a real time quantitative reporter of heterologous protein production. Biotechnol. Prog. 14: 351–354.
Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC (1994) Green fluorescent protein as a marker for gene expression. Science 263: 802–805.
Côté J, Bourget L, Garnier A, Kamen A (1997) Study of adenovirus production in serum-free 293SF suspension culture by GFPexpression monitoring [ln Process Citation]. Biotechnol. Prog. 13: 709–714.
Cubitt AB, Heim R, Adams SR, Boyd AE, Gross LA, Tsien RY (1995) Understanding, improving and using green fluorescent proteins. Trends Biochem. Sci. 20: 448–455.
DeLisa MP, Li J, Rao G, Weigand WA, Bentley WE (1999) Monitoring GFP-operon fusion protein expression during high cell density cultivation of Escherichia coli using an on-line sensor. Biotech. Bioeng. 65: 54–64.
Gerdes HH, Kaether C (1996) Green fluorescent protein: applications in cell biology. FEBS Lett. 389: 44–47.
Heim R, Cubitt AB, Tsien RY (1995) Improved green fluorescence. Nature 373: 663–664.
Hunt L, Jordan M, De Jesus M, Wurm FM (1999) GFP-expressing mammalian cells for fast, sensitive, noninvasive cell growth assessment in a kinetic mode. Biotechnol. Bioeng. 65: 201–205.
Inouye S, Tsuji FI (1994) Aequorea green fluorescent protein. Expression of the gene and fluorescence characteristics of the recombinant protein. FEBS Lett. 341: 277–280.
Lamarche N, Massie B, Richer M, Paradis H, Langelier Y (1990) High level expression in 293 cells of the herpes simplex virus type 2 ribonucleotide reductase subunit 2 using an adenovirus vector. J. Gen. Virol. 71: 1785–1792.
Massie B, Mosser D, Koutroumanis M, Vitte-Mony I, Lamoureux L, Couture F, Paquet L, Guilbault C, Dionne J, Chahla D, Jolicoeur P, Langelier Y (1998) New adenovirus vectors for protein production and gene transfer. Cytotechnology 28: 53–65.
Prasher DC, Eckenrode VK, Ward WW, Prendergast FG, Cormier MJ (1992) Primary structure of the Aequorea victoria green-fluorescent protein. Gene 111: 229–233.
Randers-Eichhorn L, Albano CR, Sipior J, Bentley WE, Rao G (1997) On-line green fluorescent protein sensor with LED excitation. Biotechnol. Bioeng. 55: 921–926.
Stauber RH, Horie K, Carney P, Hudson EA, Tarasova NI, Gaitanaris GA, Pavlakis GN (1998) Development and applications of enhanced green fluorescent protein mutants. Biotechniques 24: 462–471.
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Gilbert, PA., Garnier, A., Jacob, D. et al. On-line measurement of green fluorescent protein (GFP) fluorescence for the monitoring of recombinant adenovirus production. Biotechnology Letters 22, 561–567 (2000). https://doi.org/10.1023/A:1005672410905
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DOI: https://doi.org/10.1023/A:1005672410905