Optimization of Insect Cell Based Protein Production Processes - Online Monitoring, Expression Systems, Scale Up

Druzinec, Damir; Salzig, Denise; Brix, Alexander; Kraume, Matthias; Vilcinskas, Andreas; Kollewe, Christian; Czermak, Peter

doi:10.1007/10_2013_205

Cited by 15 publications

(16 citation statements)

References 128 publications

(119 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The FDA has addressed this issue by launching the process analytical technology (PAT) initiative [76], whereas the European Medicines Agency (EMA) has implemented the similar concept of continuous process verification [77]. To fulfil these demanding requirements, bioreactor systems have been fitted with diverse monitoring systems including near infrared spectroscopy, and in situ microscopy [78][79][80]. Multivariate data analysis is then applied to determine a benchmark known as the "golden batch".…”

Section: Process Controlmentioning

confidence: 99%

Concepts for the Production of Viruses and Viral Vectors in Cell Cultures

Grein¹,

Weidner²,

Czermak³

2017

New Insights Into Cell Culture Technology

Self Cite

View full text Add to dashboard Cite

The industrial-scale manufacturing of viruses or virus-like particles in cell culture is necessary for gene therapy and the treatment of cancer with oncolytic viruses. Complex multistep processes are required in both cases, but the low virus titers in batch cultures and the temperature sensitivity of the virus particles limit the production scale. To meet commercial and regulatory requirements, each process must be scalable and reproducible and must yield high virus titers. These requirements are met by establishing a cell culture process that matches the properties of the virus/host-cell system and by using serum-free cell culture medium. This chapter focuses on two case studies to consider the different aspects of process design, such as the reactor configuration and operational mode: the continuous production of retroviral pseudotype vectors in a retroviral packaging cell line and the production of oncolytic measles virus vectors for cancer therapy.

show abstract

Section: Process Controlmentioning

confidence: 99%

Concepts for the Production of Viruses and Viral Vectors in Cell Cultures

Grein¹,

Weidner²,

Czermak³

2017

New Insights Into Cell Culture Technology

Self Cite

View full text Add to dashboard Cite

show abstract

“…During a virus production process, the radius/volume of the host cell changes due to the infection process (Druzinec et al, 2013;Emma & Kamen, 2012;Justice et al, 2011;Negrete et al, 2007). By applying DS, the cell radius r can be calculated using the critical frequency fc, considering the intracellular and extracellular conductivities, σ i and σ e (Petiot et al, 2010;Schwan, 1957).…”

Section: Discussionmentioning

confidence: 99%

“…It is widely used in yeast fermentation (Asami, Gheorghiu, & Yonezawa, 1999;Huang, Holzel, Pethig, & Xiao, 1992) and has also been used to monitor the expansion of animal cells, the differentiation of stem cells, and virus-based production processes (Downey, Graham, Breit, & Glutting, 2014;Druzinec et al, 2013;Justice et al, 2011;Negrete, Esteban, & Kotin, 2007). It is widely used in yeast fermentation (Asami, Gheorghiu, & Yonezawa, 1999;Huang, Holzel, Pethig, & Xiao, 1992) and has also been used to monitor the expansion of animal cells, the differentiation of stem cells, and virus-based production processes (Downey, Graham, Breit, & Glutting, 2014;Druzinec et al, 2013;Justice et al, 2011;Negrete, Esteban, & Kotin, 2007).…”

Section: Several Other Viruses Have Been Investigated As Virotherapymentioning

confidence: 99%

High titer oncolytic measles virus production process by integration of dielectric spectroscopy as online monitoring system

Grein

Loewe

Dieken

et al. 2018

Biotech & Bioengineering

Self Cite

View full text Add to dashboard Cite

Oncolytic viruses offer new hope to millions of patients with incurable cancer. One promising class of oncolytic viruses is Measles virus, but its broad administration to cancer patients is currently hampered by the inability to produce the large amounts of virus needed for treatment (10 -10 virus particles per dose). Measles virus is unstable, leading to very low virus titers during production. The time of infection and time of harvest are therefore critical parameters in a Measles virus production process, and their optimization requires an accurate online monitoring system. We integrated a probe based on dielectric spectroscopy (DS) into a stirred tank reactor to characterize the Measles virus production process in adherent growing Vero cells. We found that DS could be used to monitor cell adhesion on the microcarrier and that the optimal virus harvest time correlated with the global maximum permittivity signal. In 16 independent bioreactor runs, the maximum Measles virus titer was achieved approximately 40 hr after the permittivity maximum. Compared to an uncontrolled Measles virus production process, the integration of DS increased the maximum virus concentration by more than three orders of magnitude. This was sufficient to achieve an active Measles virus concentration of > 10 TCID ml .

show abstract

“…There are several cell lines like this available such as stable transfected Sf9, as well as the Drosophila melanogaster cell lines like Schneider 2 (S2) and 3 (S3) . The stable transfected cells can also grow adherent in a monolayer or as a suspension in serum‐ or protein‐free media, which enables large‐scale cultivation and production in a bioreactor …”

Section: Heterologous Expression Hosts For Insect‐derived Ampsmentioning

confidence: 99%

“…23 The stable transfected cells can also grow adherent in a monolayer or as a suspension in serum-or protein-free media, which enables large-scale cultivation and production in a bioreactor. 24 Large-scale production for both insect expression systems have been executed with varying success. b-galactosidase was expressed 1,0003 lower in stable cell lines than in the BEVS.…”

Section: Insect Cells-a True Alternative For the Expression Of Insectmentioning

confidence: 99%

Considerations for the process development of insect‐derived antimicrobial peptide production

Müller

Salzig

Czermak

2014

Biotechnology Progress

Self Cite

View full text Add to dashboard Cite

Antimicrobial peptides (AMPs) could evolve into new therapeutic lead molecules against multi-resistant bacteria. As insects are a rich source of AMP, the identification and characterization of insect-derived AMPs is particularly emphasized. One challenge of bringing these molecules into market, e.g., as a drug, is to develop a cost-efficient large-scale production process. Due to the fact that a direct AMP isolation from insects is not economical and that chemical synthesis is recommended for peptide sizes below 40 amino acids, a viable option is heterologous AMP production. Therefore, previous knowledge concerning the expression of larger proteins can be adapted, but due to the AMP nature (e.g., small size, bactericide) additional challenges have to be faced during up and downstream processing. Nonetheless the bottleneck for large-scale AMP production is the same as for proteins; mainly the downstream process. This review introduces opportunities for insect-derived AMP production, like the choice of the expression system (based on previously derived data), depending on the AMP nature, as well as new purification strategies like elastin-like peptide/intein based purification strategies. All of these aspects are discussed with regard to large-scale processes and costs.

show abstract

Optimization of Insect Cell Based Protein Production Processes - Online Monitoring, Expression Systems, Scale Up

Cited by 15 publications

References 128 publications

Concepts for the Production of Viruses and Viral Vectors in Cell Cultures

Concepts for the Production of Viruses and Viral Vectors in Cell Cultures

High titer oncolytic measles virus production process by integration of dielectric spectroscopy as online monitoring system

Considerations for the process development of insect‐derived antimicrobial peptide production

Contact Info

Product

Resources

About