Key Parameters of Measles Virus Production for Oncolytic Virotherapy

Weiss, Katja; Salzig, Denise; Mühlebach, Michael D.; Cichutek, Klaus; Pörtner, Ralf; Czermak, Peter

doi:10.3844/ajbbsp.2012.81.98

Cited by 16 publications

(3 citation statements)

References 60 publications

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“…While these virus particles are stabilized with different agents or are even lyophilized, this stability studies are not representative for measles virus particles under production conditions. The key parameters for MV production for the use in cancer therapy were already discussed in a previous review (Weiss et al, 2012).…”

Section: Ajbbmentioning

confidence: 99%

Influence of Process Conditions on Measles Virus Stability

Weiss

Salzig

Röder

et al. 2013

American Journal of Biochemistry and Biotechnology

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Recombinant measles viruses are currently tested in clinical trials as oncolytic agent to be applied in cancer therapy. Contrary to their use as vaccine where 10 3 infectious virus particles per dose are needed, for cancer therapy 10 9 virus particles should be provided per dose. This leads to other challenges for the production process when compared to vaccine production. This study presents measles virus stability with regard to conditions during production and storage of the virus. Relevant process parameters such as temperature (4-37°C), pH (pH 4-11), conductivity (1.5 to 137.5 mS cm ) and oxygen partial pressure were analyzed. The infectivity of measles virus particles decreased highly at 37 and 32°C, while at 22 and 4°C it remained stable for several hours or even days, respectively. The thermal inactivation reactions followed first order kinetics and the thermodynamic parameters enthalpy and entropy were estimated. Towards changes in pH measles virus particles were very sensitive, while no inactivation could be observed with varying conductivity. Measles virus incubation at an oxygen partial pressure of 100% did not lead to any loss of infectivity. The results show which parameters should be considered and controlled strongly in the production process to further raise measles virus yields for the high amount needed in cancer therapy approaches.

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Section: Ajbbmentioning

confidence: 99%

Influence of Process Conditions on Measles Virus Stability

Weiss

Salzig

Röder

et al. 2013

American Journal of Biochemistry and Biotechnology

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“…Measles virus (MV) is a leading candidate owing to the excellent safety profile of the measles vaccine [2]. However, at least 10 8 infectious MV particles are needed per dose for cancer therapy [3,4] and this has prompted extensive studies focusing on the optimization of upstream processes [5][6][7][8][9][10]. The deeper understanding of MV production gained from these studies has shown that shear stress and aeration play a key role in process productivity [7].…”

Section: Introductionmentioning

confidence: 99%

Tangential Flow Filtration for the Concentration of Oncolytic Measles Virus: The Influence of Filter Properties and the Cell Culture Medium

et al. 2019

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The therapeutic use of oncolytic measles virus (MV) for cancer treatment requires >108 infectious MV particles per dose in a highly pure form. The concentration/purification of viruses is typically achieved by tangential flow filtration (TFF) but the efficiency of this process for the preparation of MV has not been tested in detail. We therefore investigated the influence of membrane material, feed composition, and pore size or molecular weight cut-off (MWCO) on the recovery of MV by TFF in concentration mode. We achieved the recovery of infectious MV particles using membranes with a MWCO ≤ 300 kDa regardless of the membrane material and whether or not serum was present in the feed. However, serum proteins in the medium affected membrane flux and promoted fouling. The severity of fouling was dependent on the membrane material, with the cellulose-based membrane showing the lowest susceptibility. We found that impurities such as proteins and host cell DNA were best depleted using membranes with a MWCO ≥ 300 kDa. We conclude that TFF in concentration mode is a robust unit operation to concentrate infectious MV particles while depleting impurities such as non-infectious MV particles, proteins, and host cell DNA.

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“…In the first process type, cells are used to produce a therapeutic virus. Vero cells were used as an example for adherent cells used for therapeutic virus production (Weiss et al, 2012; where cell detachment is only crucial during the seeding train to achieve enough cell mass for virus infection. MA104 cells are adherent cells also used in vaccine production and were chosen in the current study as they are very adhesive.…”

Section: Introductionmentioning

confidence: 99%

CELL DETACHMENT BY PROLYL-SPECIFIC ENDOPEPTIDASE FROM <i>WOLFIPORIA COCOS</i>

Mika

Cierpka

Lange

et al. 2014

American Journal of Biochemistry and Biotechnology

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As requirements for Advanced Therapy Medicinal Product (ATMP) production differ from other production processes (e.g., therapeutic protein production), cell detachment is often a crucial step for the process success. In most cases, cell detachment is done enzymatically. Although many peptidases are established in cell culture in R&D, e.g., Trypsin as gold standard, many of them seem to be unsuitable in ATMP production processes. Therefore, the present study investigated a novel endopeptidase used in food biotechnology for its applicability in ATMP processes where cell detachment is needed. The Prolyl-specific Peptidase (PsP) is of nonmammalian origin and considered as safe for humans. PsP was purified from the supernatant of the fungus Wolfiporia cocos. The isolation and purification resulted in an enzyme solution with 0.19 U mg −1 prolylspecific activity. By in silico analysis it was confirmed that attachment-promoting proteins can be cleaved by PsP in a similar amount than with Trypsin. Further the proteolytic activity was determined for PsP and Trypsin by using the same enzymatic assay. Detachment with both enzymes was compared for cells used in typical therapeutic production processes namely a mesenchymal stem cell line (hMSC-TERT) as a model for a cell therapeutic, Vero and MA104 cells used for viral therapeutic or vaccine production. The cell detachment experiments were performed with comparable enzyme activities (1.6 U mL −1). hMSC-TERT detachment was faster with PsP than with Trypsin. For Vero cells the detachment with PsP was not only faster but also more efficient. For MA104 cells the detachment rate with PsP was similar to Trypsin. For all cell types, detachment with PsP showed less influence on cell growth and metabolism compared to standard Trypsin.Thus, three cell types used in ATMP, viral therapeutics or vaccine production can be detached efficiently and gently with PsP. Therefore, PsP shows potential for cell detachment in ATMP and viral/vaccine production processes.

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Key Parameters of Measles Virus Production for Oncolytic Virotherapy

Cited by 16 publications

References 60 publications

Influence of Process Conditions on Measles Virus Stability

Influence of Process Conditions on Measles Virus Stability

Tangential Flow Filtration for the Concentration of Oncolytic Measles Virus: The Influence of Filter Properties and the Cell Culture Medium

CELL DETACHMENT BY PROLYL-SPECIFIC ENDOPEPTIDASE FROM <i>WOLFIPORIA COCOS</i>

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