Improved Production of Recombinant Proteins by the Baculovirus Expression System

Radford, Kathryn M.; Reid, Steve; Greenfield, P. F.

doi:10.1007/978-94-011-2844-5_57

Cited by 9 publications

(14 citation statements)

References 3 publications

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“…In the large-scale production of recombinant proteins, cultures infected at high-cell densities produce higher volumetric protein levels than those infected at low cell densities (Wong et al, 1996;Yang et al, 1996). However, high-cell density infections are limited by the nutrient supply from the medium (Radford et al, 1992;Radford et al, 1997b;Zhang et al, 1994). Recently, Tsao et al (1996) studied the production of a parvovirus B19 vaccine in insect cells coinfected with two baculoviruses.…”

Section: Discussionmentioning

confidence: 99%

“…The production of recombinant proteins, using Sf9 cells in shaker-or bioreactor-suspension cultures with the SF900 II medium have been studied extensively to date, using ␤-Galactosidase (␤-Gal) as a model protein product (Power et al, 1992;Radford et al, 1992;1997a;Wong et al, 1996). This previous work with ␤-Gal has established that maximum volumetric yields using the Sf9/SF900 II /rAcNPV system in batch culture are achieved when cells are infected, (at a high MOI, multiplicity of infection), at around 4 × 10 6 cells/mL (Wong et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

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Optimized production of recombinant bluetongue core-like particles produced by the baculovirus expression system

Zheng

Greenfield

Reid

1999

Biotechnol. Bioeng.

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The baculovirus‐expression vector system (BEVS) has been widely used for the experimental production of many human and animal single‐ and multi‐unit vaccines, heterologous proteins, and viral insecticides. In this work, the production of recombinant bluetongue virus core‐like particles (CLPs), using Sf9 cells in shaker‐suspension culture with the SF900 II medium (GIBCO, NY), has been studied. This system involved the simultaneous production of two proteins, VP7 and VP3, and was shown to achieve high volumetric productivities. The key parameters of the time of infection (TOI), and the multiplicity of infection (MOI) were studied. The results show that the peak‐volumetric yields and cell‐specific yields achieved using low MOIs at low‐cell densities were the same as those obtained following infections with a high MOI at high‐cell densities. This work establishes the feasibility of using low MOIs in the baculovirus system to produce complex multiprotein particles. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 65: 600–604, 1999.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Optimized production of recombinant bluetongue core-like particles produced by the baculovirus expression system

Zheng

Greenfield

Reid

1999

Biotechnol. Bioeng.

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“…The estimated specific rates of production for IPL-41 and Sf90011 were 10.3 (22.5) and 9.8 (21.5) PFU per cell per hour, respectively. The estimated times of commencement in the two media were 22 (k2) and 17 (22) hours postinfection, respectively, and release was estimated to continue until 76…”

Section: Extracellular Virus: K T~~~ T~~~ and Avpmentioning

confidence: 99%

“…The product yield decreases sharply when cultures are infected later than a certain optimal TOI.279, 22,26 This is the basis for the rule of thumb that cells must be infected in the early to midexponential phase of a culture. We and others have observed that high yields can be maintained even in the late exponential phase, if the cells are resuspended in fresh medium.z3'4'zz~23 Hence, production appears limited either by depletion of nutrients or accumulation of inhibitors.…”

Section: Introductionmentioning

confidence: 98%

Modeling and optimization of the baculovirus expression vector system in batch suspension culture

Power

Reid

Radford

et al. 1994

Biotech & Bioengineering

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A mathematical model has been developed that predicts the cell population dynamics and production of recombinant protein and infective extracellular virus progeny by insect cells after infection with baculovirus in batch suspension culture. Infection in the model is based on the rate of virus attachment to suspended insect cells under culture conditions. The model links the events following infection with the sequence of gene expression in the baculovirus replicative cycle. Substrate depletion is used to account for the decrease in product yield observed when infecting at high cell densities. Model parameters were determined in shaker flasks for two media: serum-supplemented IPL-41 medium and serum free Sf900II medium. There was good agreement between model predictions and the results from an independent series of experiments performed to validate the mode. The model predicted: (1) the optimal time of infection at high multiplicity of infection: (2) the timing and magnitude of recombinant protein production in a 2-L bioreactor; and (3) the timing and magnitude of recombinant protein production at multiplicities of infection from 0.01 to 100 plaque-forming units per cell. Through its ability to predict optimal infection strategies in batch suspension culture, the model has use in the design and optimization of large-scale systems for the production of recombinant products using the baculovirus expression vector system.

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“…IO, No. 6 et al, 1990Kool et al, 1991;Licari and Bailey, 1991;Radford et al, 1992;Wickham et al, 1991;Zhang et al, 1993). Nevertheless, the possibility of generalizing the effect of each major variable on expression levels is unlikely because most of the previous work on identifylng such factors has been limited to the AcNPV/Sf9 expression system and different results have been reported.…”

Section: Introductionmentioning

confidence: 99%

Optimum Infection Conditions for Recombinant Protein Production in Insect Cell (Bm5) Suspension Culture

Zhang

Kalogerakis

Behie

et al. 1994

Biotechnology Progress

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The baculovirus/insect cell expression system is an efficient and practical method for the production of many active therapeutic proteins on a large scale. The advantages of suspension cultures have been demonstrated with the study of a baculovirus/insect cell (BmNPV/Bm5) expression system for the production of recombinant chloramphenicol acetyltransferase (CAT), a model heterologous protein. Key infection parameters such as infection time and multiplicity of infection were examined systematically for the maximization of protein production. Furthermore, emphasis was placed on the development of possible medium replenishment strategies, which were necessary to achieve higher volumetric protein production from the infection of high-density cell cultures without sacrificing specific protein productivity. The highest protein production was achieved with the infection of suspended cells in the mid to late exponential growth phase.

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Improved Production of Recombinant Proteins by the Baculovirus Expression System

Cited by 9 publications

References 3 publications

Optimized production of recombinant bluetongue core-like particles produced by the baculovirus expression system

Optimized production of recombinant bluetongue core-like particles produced by the baculovirus expression system

Modeling and optimization of the baculovirus expression vector system in batch suspension culture

Optimum Infection Conditions for Recombinant Protein Production in Insect Cell (Bm5) Suspension Culture

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