Assessing two strategies for production of murine ascites with anti-SARS-CoV-2 monoclonal antibodies

Authors

  • Joel Javier Pérez-Paz Center for Genetic Engineering and Biotechnology of Sancti Spiritus, Circunvalante Norte, Olivos 3, Sancti Spiritus, Cuba
  • Reinaldo Blanco Center for Genetic Engineering and Biotechnology of Sancti Spiritus, Circunvalante Norte, Olivos 3, Sancti Spiritus, Cuba
  • Dayamí Dorta Center for Genetic Engineering and Biotechnology of Sancti Spiritus, Circunvalante Norte, Olivos 3, Sancti Spiritus, Cuba
  • Andy Domínguez Center for Genetic Engineering and Biotechnology of Sancti Spiritus, Circunvalante Norte, Olivos 3, Sancti Spiritus, Cuba
  • Maylin Pérez-Bernal Center for Genetic Engineering and Biotechnology of Sancti Spiritus
  • Celia Tamayo Center for Genetic Engineering and Biotechnology of Sancti Spiritus, Circunvalante Norte, Olivos 3, Sancti Spiritus, Cuba
  • Carlos Hernández Center for Genetic Engineering and Biotechnology of Sancti Spiritus, Circunvalante Norte, Olivos 3, Sancti Spiritus, Cuba
  • Ricardo Pina Center for Genetic Engineering and Biotechnology of Sancti Spiritus, Circunvalante Norte, Olivos 3, Sancti Spiritus, Cuba
  • Javier Díaz Center for Genetic Engineering and Biotechnology of Sancti Spiritus, Circunvalante Norte, Olivos 3, Sancti Spiritus, Cuba
  • Shaylí Pérez Center for Genetic Engineering and Biotechnology of Sancti Spiritus, Circunvalante Norte, Olivos 3, Sancti Spiritus, Cuba
  • Ivis Pasarón Center for Genetic Engineering and Biotechnology of Sancti Spiritus, Circunvalante Norte, Olivos 3, Sancti Spiritus, Cuba
  • Enrique Pérez Center for Genetic Engineering and Biotechnology of Sancti Spiritus, Circunvalante Norte, Olivos 3, Sancti Spiritus, Cuba

DOI:

https://doi.org/10.52331/cvj.v26i3.31

Keywords:

ascites, hybridoma, mice, mineral oil

Abstract

Studies were conducted to improve the production of murine ascites with monoclonal antibodies that recognize SARS-CoV-2 proteins. BALB/c mice were primed with 0.5 mL of mineral oil into the abdominal cavity. Seven days after priming, mice were divided in two groups: the group 1 was inoculated intraperitoneally with 2x106 cells/mL of MAb-secreting hybridomas against the nucleocapsid and spike proteins of SARS-CoV-2; the group 2 was injected simultaneously with the same inoculum of hybridoma cells and mineral oil, 18 days after priming. No disturbances or suffering signals were observed in mice from both groups, suggesting that double administration of mineral oil did not produce significant distress with respect to the single dose used for priming, and that none of the hybridoma cell lines were particularly aggressive for the inoculated mice. Ascites was collected in 90.48% and 97.68% of mice from groups 1 and 2, respectively. Ascites was not collected in 7.42% of all mice. The main cause was they never developed ascites tumors but no solid tumors were observed either. The volume of ascitic fluid per mouse was increased significantly in mice from group 2, and there were no significant differences between groups in terms of the concentration of IgG in clarified ascites. According to these results, to obtain higher amounts of MAb the strategy applied in group 2 should be used, since it showed the best results in the development of ascites tumors and it significantly increased the volume of ascites fluid per mouse. This could allow the use of fewer animals for ascites production, which is an ethical and economic benefit.

References

Mattioli, I.A.; Hassan, A.; Oliveira, O.N.; Crespilho, F.N. On the Challenges for the diagnosis of SARS-CoV-2 based on a review of current methodologies. ACS Sens. 2020, 5(12), 3655–3677, doi.org/10.1021/acssensors.0c01382.

Clark, A.; Befus, D.; O'Hashi, P.; Hart, F.; Schunk, M.; Fletch, A.; Griffin, G. Guidelines on: antibody production. Canadian Council on Animal Care, 2002.

Bonistalli, K.N. Monoclonal antibody production: a comparison of in vitro and in vivo methods and their use in Clostridial vaccine manufacture. Master of Science Thesis in Veterinary Medicine, Institute of Veterinary, Animal and Biomedical Sci-ences, Massey University, New Zealand, 2013.

Nessa, M.U.; Rahman, M.A.; Kabir, Y. Plant-produced monoclonal antibody as immunotherapy for cancer. BioMed Res. Int. 2020, 3038564, doi.org/10.1155/2020/3038564.

Chiarella, P.; Fazio, V.M. Mouse monoclonal antibodies in biological research: strategies for high-throughput production. Biotechnol. Lett. 2008, 30(8), 1303-1310, doi.org/10.1007/s10529-008-9706-5.

Ward, P.; Adams, J.; Faustman, D.; Gebhart, G.; Geistfeld, J.; Imbaratto, J.; Peterson, N.; Quimby, F.; Marshak-Rothstein, A.; Rowan, A.; Scharff, M. Monoclonal Antibody Production. Institute for Laboratory Animal Research. National Research Council. National Academy Press Washington, USA, 1999.

Amyx, H.L. Control of animal pain and distress in antibody production and infectious disease studies. J. Am. Vet. Med. Assoc. 1987, 191, 1287-1289.

Jackson, L.R.; Trudel, L.J.; Fox, J.G.; Lipman, N.S. Monoclonal antibody production in murine ascites: I. Clinical and patho-logical features. Lab. Anim. Sci. 1999, 49, 70-80.

Mendoza, O.; Valdés, R.; González, M.; Alemán, R.; Álvarez, T.; Padilla, S.; Tamayo, A.; Reyes, B.; Geada, D.; Fernández, E.; Fuentes, D.; Hernández, O.; Zuasnabar, L. Influence of the number of animals on the production of monoclonal antibody CB.Hep-1 by the Ascites method. Biotecnol. Apl. 2009, 26(2), 133-137.

Peterson, N.C. Behavioral, clinical, and physiological analysis of mice used for ascites monoclonal antibody production. Comp. Med. 2000, 50(5), 516-526.

Institutional Animal Care and Use Committee (IACUC), UTA Monoclonal Antibodies/Mouse Ascites Sop (V1.0) University of Texas at Arlington, USA, 2016.

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Published

2021-12-31

How to Cite

Pérez-Paz, J. J. ., Blanco, R., Dorta, D. ., Domínguez, A. ., Maylin Pérez-Bernal, Tamayo, C. ., Hernández, C. ., Pina, R. ., Díaz, J. ., Pérez, S. ., Pasarón, I. . and Pérez, E. . (2021) “Assessing two strategies for production of murine ascites with anti-SARS-CoV-2 monoclonal antibodies”, Cluj Veterinary Journal, 26(3), pp. 2–7. doi: 10.52331/cvj.v26i3.31.