Inactivation of foot and mouth disease virus for vaccine production

Inactivation is the loss by a virus of ability to reproduce and infect susceptible animals while retaining its antigenic properties. In this paper, the effectiveness of aminoethylethanolamine when used as an FMDV inactivant is shown. The inactivation rates under selected parameters, effect of aminoethylethanolamine on virus stability during inactivation and on vaccine immunogenicity after storage at 2–8 °С were determined. The method for calculation of 50% aminoethylethanolamine inactivating concentration (IC₅₀) which enables to determine quality parameters of the virus-containing suspension, to compare the inactivating agent activities and their ability to ensure the vaccine innocuity within the given period of time is presented. It was established that IC₅₀ for purified and non-purified virus-containing suspensions was identical (0.0045%), and its safety after 12 h of inactivation was one TCID₅₀ per 10⁹–10¹¹ L of the virus containing suspension. It was also found that double increase in inactivation time increased the virucidal activity of aminoethylethanolamine by a factor of 1.8 for serotype O and 2.4 for serotype A. At the same time, the removal of cell debris had no significant effect on the inactivation process. Aminoethylethanolamine does not destroy 146S virus particles and it was confirmed by immunogenicity testing of the vaccines during storage. This means that 15% aqueous solution of aminoethylethanolamine, manufactured by Russian Company OOO “BIOKHIMRESURS” (Vladimir) complies with high quality standards. Immunogenicity test of bivalent FMD vaccine for cattle by challenging demonstrated that its potency was 10.08 protective doses per 2 cm³ of the vaccination dose.

1. Sergeev V. A., Nepoklonov E. A., Aliper T. I. Viruses and viral vaccines. Moscow: Biblionika; 2007. 524 p. (in Russ.)

2. Vallée H., Carré H., Rinjard P. Sur l’immunisation anti-aphteuse par le virus formolé. Rev. gén. Méd. vét. 1926; 35: 129–134. (in French)

3. Barteling S. J., Woortmeyer R. Formaldehyde inactivation of foot-and-mouth disease virus. Conditions for the preparation of safe vaccine. Arch. Virol. 1984; 80 (2–3): 103–117. DOI: 10.1007/BF01310652.

4. Gard S. Theoretical considerations in the inactivation of viruses by chemical means. Ann. N. Y. Acad. Sci. 1960; 83: 638–648. DOI: 10.1111/j.1749-6632.1960.tb40934.x.

5. Melvin P. A., Pryce J.-M. G. Production of viral antigens with N-acetyl-ethyleneimine. Patent No. US3318775A United States, Int. C12N7/00. Imperial Chemical Industries Ltd. Date of filing: 29.01.1963. Date of publication: 09.05.1967. Available at: https://patents.google.com/patent/US3318775A/en.

6. Ulupov N. A., Mikhalishin V. V., Gusev A. A., Lyozova T. N. Aminoetiletilenimin – sredstvo inaktivatsii infektsionnosti virusa yashchura = Aminoethylethanolamine is an inactivating agent of foot and mouth disease virus infectivity. Sovremennye aspekty veterinarnoi patologii zhivotnykh: materialy konferentsii, posvyashchennoi 40-letiyu so dnya osnovaniya VNIIZZh (23–25 noyabrya 1998 g.) = Modern aspects of animal pathology: proceedings of the conference, devoted to 40th anniversary of ARRIAH (November 23–25, 1998). Vladimir; 1998; 53–62. (in Russ.)

7. Pay T. W. F., Telling R. C., Kitchener B. L., Southern J. Some observations of FMD virus inactivation with acetylethyleneimine. In: European Commission for the Control of Foot-and-Mouth Disease. Report of the Meeting of the Research Group of the Standing Technical Committee. Romе: FAO; 1972; 81–86. Available at: https://www.fao.org/3/ca9381en/ca9381en.pdf.

8. Brown F., Crick J. Application of agar-gel diffusion analysis to a study of the antigenic structure of inactivated vaccines prepared from the virus of foot-and-mouth disease. J. Immunol. 1959; 82 (5): 444–447. PMID: 13641679.

9. Brown F., Hyslop N. St G., Crick J., Morrow A. W. The use of acetylethyleneimine in the production of inactivated foot-and-mouth disease vaccines. J. Hyg. (Lond). 1963; 61 (3): 337–344. DOI:10.1017/s0022172400039620.

10. Bahnemann H. G. Inactivation of viral antigens for vaccine preparation with particular reference to the application of binary ethylenimine. Vaccine. 1990; 8 (4): 299–303. DOI: 10.1016/0264-410x(90)90083-x.

11. Bahnemann H. G. Binary ethylenimine as an inactivant for foot-and-mouth disease virus and its application for vaccine production. Arch. Virol. 1975; 47 (1): 47–56. DOI: 10.1007/BF01315592.

12. Mikhalishin D. V. Development of technology for manufacture of FMD emulsion vaccine: Author’s Abstract of Thesis for degree of Doctor of Science (Veterinary Medicine). Vladimir; 2021. 39 p. (in Russ.)

13. Ulupov N. А. Vliyanie bufernogo rastvora na inaktivatsiyu virusa yashchura formal’degidom = Effect of buffer solution on foot and mouth disease virus inactivation with formaldehyde. Veterinariya. 1974; 9: 38–39. (in Russ.)

14. Ulupov N. A., Dudnikov A. I. Izuchenie aktivnosti khimicheskikh preparatov pri inaktivatsii virusa yashchura = Study of chemical activity for foot and mouth disease virus inactivation. Aktual’nye problemy veterinarnoi virusologii: tezisy dokladov nauchnoi konferentsii VNIYaI = Topical issues of veterinary virology: conference paper abstracts of All-Russian FMD Research Institute scientific conference. Vladimir; 1983; 34–35. (in Russ.)

15. Ulupova T. N., Ulupov N. A., Rybakov S. S., Koropov V. N., Dudnikov A. I. Sravnitel’noe izuchenie inaktiviruyushchei aktivnosti aziridinov po otnosheniyu k virusu yashchura = Comparative study of aziridine inactivating activities in relation to foot and mouth disease virus. Aktual’nye problemy veterinarnoi virusologii: tezisy dokladov nauchnoi konferentsii VNIYaI = Topical issues of veterinary virology: conference paper abstracts of All-Russian FMD Research Institute scientific conference. Vladimir; 1983; 37–39. (in Russ.)

16. Warrington R. E., Cunliffe H. R., Bachrach H. L. Derivatives of aziridine as inactivants for foot-and-mouth disease virus vaccines. Am. J. Vet. Res. 1973; 34 (8): 1087–1091. PMID: 4356910.

17. Ulupov N. A. Evaluation technique of chemical inactivation properties. Vladimir; 1980. 4 p. (in Russ.)

18. Barteling S. J. Development and performance of inactivated vaccines against foot and mouth disease. Rev. Sci. Tech. 2002; 21 (3): 577–588. DOI: 10.20506/rst.21.3.1361.

19. Aarthi D., Ananda Rao K., Robinson R., Srinivasan V. A. Validation of binary ethyleneimine (BEI) used as an inactivant for foot and mouth disease tissue culture vaccine. Biologicals. 2004; 32 (3): 153–156. DOI: 10.1016/j.biologicals.2004.09.001.

20. Ulupov N. A., Dudnikov A. I., Mikhalishin V. V. Lapinized foot-and-mouth disease vaccine of the FGI “ARRIAH” origin. Proceedings of the Federal Centre for Animal Health. 2008; 6: 323–340. eLIBRARY ID: 14933111. (in Russ.)

21. El’kina Yu. S. Vaccines against foot and mouth disease types O, Asia-1, A to induce early immunity in animals: Author’s Abstract of Thesis for degree of Candidate of Science (Veterinary Medicine). Vladimir; 2021. 21 p. (in Russ.)

22. Eremets V. I., Bol’shаkova M. V., Fomina G. А., et al. Izyskanie effektivnykh inaktivantov pri krupnomasshtabnom proizvodstve protivoyashchurnykh vaktsin = Search for effective inactivating agents in largescale production of foot and mouth disease vaccines. Nauchnye osnovy proizvodstva veterinarnykh biologicheskikh preparatov: tezisy dokladov Vserossiiskoi nauchno-prakticheskoi konferentsii, posvyashchennoi 30-letiyu instituta (8–9 iyunya 2000 g.) = Scientific basis for production of veterinary biological products: conference paper abstracts of All-Russian Scientific and Practical Conference devoted to 30th Anniversary of the Institute (June 8–9, 2000). Shchelkovo; 2000; 52–54. (in Russ.)

23. Pigoleva I. V., Sharypov A. S., Shabalina T. N., Zaglyadova S. V., Kitova M. V., Antonov S. A., et al. Testing of emulsion vaccines based on white oils obtained as a result of hydrocatalytic processing of oilstock. Veterinary Science Today. 2017; (4): 42–48. (in Russ.)

24. Lozovoy D. A., Mikhalishin D. V., Lyozova T. N., Mikhalishin V. V., Konstantinov A. V., Starikov V. A., Borisov A. V. Laboratory studying efficacy of sorbate vaccine against heterologous strains of type A FMD virus. Veterinariya. 2016; 2: 16–19. eLIBRARY ID: 25522294. (in Russ.)

25. Foot and mouth disease (infection with foot and mouth disease virus). In: WOAH. Terrestrial Animal Health Code. 2022; Chapter 3.1.8. Available at: https://www.woah.org/fileadmin/Home/eng/Health_standards/tahm/3.01.08_FMD.pdf.

26.