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DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein

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Abstract

The transmembrane rabies virus glycoprotein (RVGP) is the main antigen of vaccine formulations used around the world to prevent rabies, the most lethal preventable infectious disease known. The objective of this work was to evaluate the potential of a bioreactor using wave-induced agitation in the initial steps of scaling up the rRVGP production process by a Drosophila melanogaster S2 cell line to produce rRVGP in sufficient quantities for immunization and characterization studies. Taking advantage of some remarkable features recognized in Drosophila S2 cells for scaling the culture process, a robust recombinant lineage (S2MtRVGPH-His) engineered by our group for the expression of rRVGP using a copper-inducible promoter was used in the bioreactor cultures. The WAVE Bioreactor was chosen because it represents an innovative approach to the cultivation of animal cells using single-use technology. For that purpose, we firstly established a procedure for culturing the S2MtRVGPH-His lineage in 100 mL Schott flasks. Using an inoculum of 5 × 105 cells/mL in culture medium (Sf900-III) induced with solution of CuSO4 (0.7 mM) and a convenient pH range (6.2–7.0), optimal parameter values such as time of induction (72 h) and temperature (28 °C) to increase rRVGP production could be defined. This procedure was reproduced in culture experiments conducted in a WAVE Bioreactor™ 2/10 using a 2 L Cellbag. The results in Schott flasks and in WAVE Bioreactor™ were very similar, yielding a maximum titer of rRVGP above of 1 mg.L−1. The immunization study showed that the rRVGP produced in the bioreactor was of high immunogenic quality.

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Acknowledgements

Monize Caiado Decarli acknowledges the Coordination for the Improvement of Higher Education Personnel (CAPES/Brazil) for her master fellowship and Daniela Matilde Correia acknowledges CAPES (Atração de Jovens Talentos—Process 064922/2014-01).

Funding

This study was funded by CNPq (National Council for Scientific and Technological Development)—grant number: 402439/2013-9.

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Authors and Affiliations

  1. Departament of Chemical Engineering, Federal University of São Carlos, São Carlos, SP, 13565-905, Brazil

    Monize Caiado Decarli, Diogo Peres dos Santos, Daniela Matilde Correia, Juliana de Sá da Silva & Claudio Alberto Torres Suazo

  2. Laboratory of Viral Immunology, Butantan Institute, São Paulo, SP, 05503-900, Brazil

    Renato Mancini Astray, Daniella Cristina Ventini-Monteiro, Soraia Attie Calil Jorge & Carlos Augusto Pereira

  3. Departament of Veterinary Hygiene and Public Health, São Paulo State University, Botucatu, SP, 18618-970, Brazil

    Mayra Pereira Rocca, Hélio Langoni & Benedito Donizete Menozzi

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  1. Monize Caiado Decarli
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  2. Diogo Peres dos Santos
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  3. Renato Mancini Astray
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  11. Carlos Augusto Pereira
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  12. Claudio Alberto Torres Suazo
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Correspondence to Monize Caiado Decarli.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Decarli, M.C., dos Santos, D.P., Astray, R.M. et al. DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein. Appl Microbiol Biotechnol 102, 4773–4783 (2018). https://doi.org/10.1007/s00253-018-8962-0

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  • DOI: https://doi.org/10.1007/s00253-018-8962-0

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