Production of SARS-CoV-2 pseudotyped viral particles for the evaluation of new therapies against COVID-19

Authors

  • Lucas ETCHEVERS
  • Paula SILVESTRINI
  • Natalia VELAZQUEZ
  • María Eugenia BARAVALLE
  • María Sol RENNA
  • Florencia Martín OLMOS
  • Hugo H. ORTEGA
  • Belkis MARELLI

DOI:

https://doi.org/10.14409/favecv.2021.Suppl..44-50

Keywords:

polyethyleneimine, transient transfection, lentivirus, Spike

Abstract

Passive immunotherapy as a treatment against SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2) requires to quantify neutralizing capacity of antibodies for development of effective therapies. However, the transfection reagents necessary for its production are expensive and, consequently, it is a limitation for the scaling up production. Using of polyethyleneimine (PEI), a low-cost cationic polymer, allows to avoid this. The objective of this work was to optimize a low-cost transient transfection assay to produce pseudoparticles that express the Spike protein of SARS-CoV-2. Transfection assays were performed in HEK-293T cells treated with different PEI concentrations and evaluating different incubation times. Transfection was assesed by epifluorescence microscopy and flow cytometry, showing that the transfection condition that tends to yield better results was after 48 hours of incubation using a 1/10 ratio (pDNA / PEI). In conclusion, it has been possible to adapt a protocol to produce spike-expressing lentiviral particles using a low-cost transfection reagent. This would allow to scale up the production and, following, to design a platform for evaluation of COVID-19 therapies.

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Published

2022-05-24

How to Cite

ETCHEVERS, L., SILVESTRINI, P., VELAZQUEZ, N., BARAVALLE, M. E., RENNA, M. S., OLMOS, F. M., ORTEGA, H. H., & MARELLI, B. (2022). Production of SARS-CoV-2 pseudotyped viral particles for the evaluation of new therapies against COVID-19. FAVE Sección Ciencias Veterinarias, 20(Suppl.), 44–50. https://doi.org/10.14409/favecv.2021.Suppl.44-50

Issue

Vol. 20 No. Suppl. (2021): FAVE Sección Ciencias Veterinarias - Supplement

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Original articles

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