Infectious Activity of L. Pasteur Rabies Virus Vaccine Strain Frozen-Dried in Various Protective Media And Then Stored at Various Temperatures

Viktoriia V. Varianytsia, Igor P. Vysekantsev


In this research, we studied the survival of industrial L. Pasteur rabies virus vaccine strain  after freeze-drying in various protective media and further storage at 5, –20 and –80°C. After 24-month storage (the observation period), the highest rates of the virus infectious activity were found after storage at –80°C in all the media. The maximum virus survival was found in the DMEM-based protective medium, supplemented with 0.5% bovine serum albumin, 1% gelatin and 5% sucrose. The infectious activity of the specimens, frozen-dried in the media, containing either 5% sucrose or 3% gelatin and 5% sucrose, or 10% sucrose was significantly lower. The supplement of 3% gelatin to a protective medium was established to significantly increase the time of preparations dissolution as compared to other media and the regulation standard (i. e., not more than 1 min), that could complicate their practical use. The time of specimens’ dissolution with 1% gelatin and 5% sucrose or 10% sucrose was significantly longer if comaring to those with 5% sucrose, but complied with the production regulations


Probl Cryobiol Cryomed 2018; 28(4): 333-342


rabies virus; freeze-drying; cell culture; protective media; long-term storage; virus safety; infectious activity of virus

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ATCC. Virology guide. Tips and techniques for propagating virus in tissue culture and embryonated chicken eggs. [Internet]. Manassas: ATCC; 2016. 32 p. [Cited 25.09.18] Available from:

Burkova VV, Vysekantsev IP, Lavrik AA. [Preservation of infectious activity of rabies virus industrial strains stored at various temperatures]. Zhivye I Biokosnye Sist. [Internet]. 2014;(9):1–11. [Cited 19.09.18] Available from: Russian.

Gould EA. Methods for long-term virus preservation. Molecular Biotechnology. 1999; 13(1):57–66.


Gruzdev LK, Ulasov VI, Gruzdev KN. [Estimation of the effectiveness of stabilizing media in the manufacture of a standard sample of the CVS strain of a fixed rabies virus]. Vetpatologiya. 2007; (4):32–7. Russian.

Johnson FB. Transport of viral specimens. Clinical Microbiology Reviews. 1990; 3(2):120–31.



Lalošević D, Lalošević V, Lazarević-Ivanc LJ, Knežević I. BHK-21 cell culture rabies vaccine: immunogenicity of a candidate vaccine for humans. Dev Biol. 2008; 131:421–9. PubMed

Lalosević D, Stankov S, Lazarević-Ivanc L, Lalosević V, Knezević I. Immunogenicity of BHK-rabies vaccine in human volunteers. Medicinski Pregled. 1998; 51:17–9. PubMed

Lvov DK, editor. [Medical virology]. Moscow: MIA; 2008. 656 p. Russian.

Malenovská H. The influence of stabilizers and rates of freezing рите лишние on preserving of structurally different animal viruses during скобки. lyophilization and subsequent storage. Journal of Applied Microbiology. 2014; 117(6):1810–9. CrossRef PubMed

McAleer WJ, Markus HZ, McLean AA, et al. Stability on storage at various temperatures of live measles, mumps and rubella virus vaccines in new stabilizer. Journal of Biological Standardization. 1980; 8(4):281–7.

CrossRef PubMed

Meslin FX, Kaplan MM, Koprowski H, editors. Laboratory techniques in rabies. 4th ed.. Geneva: WHO; 1996. 446 p.

Phillips GO, Harrop R, Wedlock DJ, et al. [A study of water binding in lyophilired viral vaccine systems]. Cryobiology. 1981;18(4):414–9.


Polupan IM. [Estimation of different storage methods of rabies virus vaccine strains]. Veterinarna BIotehnologIya. 2011; (20): 127–33. Ukrainian.

OIE. Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. [Internet]. Paris: World Organisation for Animal Health; 2018. Chapter 2.1.17, Rabies; p. 1–28. [Cited 25.09.18] Available from:

Rokickij PF. [Biological statistics]. Minsk: Vyisshaya shkola; 1973. 320 p. Russian.

Starodubova ES, Preobrazhenskaya OV, Kuzmenko YuV. [Vaccines against rabies: current status and development prospects]. Molekulyarnaya Biologiya. 2015; 49(4):577-84. Russian. CrossRef

Tanaka T, Takeda T, Miyajama R. Cryoprotective effect of saccharides on denaturation of catalase during freeze-drying. Chem Pharm Bull. 1991; 675(5):1091–4.


Varianytsia VV, Vysekantsev IP. Storage methods of complex RNA viruses. Probl Cryobiol Cryomed. 2017; 27(4): 287–95.


WHO Expert Committee on Biological Standardization. Recommendations for inactivated rabies vaccine for human use produced in cell substrates and embryonated eggs. In: WHO Technical Report Series – 941. Fifty-sixth report. Geneva: WHO; 2007. p. 83–132.



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