Purification of antigens from bovine alphaherpesvirus biotype 1 strain TK-A(VIEV)-b2

УДК 619:578.825.15:578.74:577.112.083
DOI 10.33861/2071-8020-2025-3-3-7

Original Empirical Research

Mukhammadiev Rish.S., Karimullina I.G., Sorokina D.A., Yarullin A.I., Khaertynov K.S., Galeeva A.G., Mukhammadiev Rin.S., Khafizova A.M.

Abstract. Infections caused by bovine alphaherpesvirus type 1 still remain a significant problem of intensive cattle farming. In this regard, the effectiveness of anti-epizootic measures is due to the timely detection of an infected animal, the correct choice of means and approaches to disease prevention. This work is devoted to the purification of antigens from bovine alphaherpesvirus biotype 1 strain TK-A(VIEV)-B2. The optimal scheme for the isolation and purification of herpesvirus antigens included disintegration of the virus-containing culture fluid by ultrasound, fractionation with polyethylene glycol 6000 reagent and purification using a step gradient of sucrose density by ultracentrifugation. As a result of the last stage of purification, we obtained seven antigen preparations. The highest serological activity, according to the data of the indirect variant of the enzyme immunoassay, was demonstrated by antigen preparations № 1.1.2, № 4.1 and № 5: the 0D450 values at a dilution of positive serum of 1:200 were 1.586±0.036 OE (K+cattle/K-cattle = 5.27), 1.081±0.023 OE (K+cattle/K-cattle = 8.72) and 1.236±0.029 OE (K+cattle/K-cattle = 4.61), respectively. Preparation № 4.1 was promising, which, according to the results of electrophoresis and immunoblotting, contained major regions of polypeptides with a molecular weight of 50 to 100 kDa, possessing antigenic properties. The antibody titer determined in the enzyme immunoassay on the 14th day after immunization with antigen preparations № 1.1.2, № 4.1 and № 5 in the blood serum of rabbits was 1:200, 1:400 and 1:400, respectively. Further studies open up prospects for using the alphaherpesvirus antigens we obtained as immunizing agents for obtaining immunoglobulin preparations with high activity and components of the enzyme immunoassay test system for serological diagnostics of infectious bovine rhinotracheitis. Keywords: bovine alphaherpesvirus biotype 1, antigen purification, polyethylene glycol fractionation, sucrose step density gradient, enzyme immunoassay test system, diagnostics, infectious rhinotracheitis, cattle breeding.

Author affiliation:

Karimullina Ilsiyar G., Ph.D. in Biology, Senior Research Fellow, Department of Virological and Ultrastructural Research, Federal Center for Toxicological, Radiation and Biological Safety; Nauchnyy gorodok-2 st., Kazan, 420075; phone: 8-904-7699225; e-mail: 89047699225@mail.ru.

Yarullin Ainur I., Ph.D. in Veterinary Sciences, Leading Research Fellow, Department of Virological and Ultrastructural Research, Federal Center for Toxicological, Radiation and Biological Safety; Nauchnyy gorodok-2 st., Kazan, 420075; phone: 8-905-3174170; e-mail: abii@mail.ru.

Sorokina Diana A., Junior Research Fellow, Department of Virological and Ultrastructural Research, Federal Center for Toxicological, Radiation and Biological Safety; st. Scientific town-2, Kazan, 420075; phone: 8-927-8875729; e-mail: diana-sorokina2013@mail.ru.

Khaertynov Kamil S., PhD in Biology, Leading Research Fellow, Department of Biochemistry and Genetic Analysis, Federal Center for Toxicological, Radiation and Biological Safety; st. Scientific town-2, Kazan, 420075; phone: 8-987-4078770; e-mail: khaerkamil@mail.ru.

Galeeva Antonina G., PhD in Veterinary Sciences, Leading Research Fellow, Department of Virological and Ultrastructural Research, Federal Center for Toxicological, Radiation and Biological Safety; Nauchnyy gorodok-2 st., Kazan, 420075; phone: 8-987-2338616; e-mail: antonina-95@yandex.ru

Mukhammadiev Rinat S., Ph.D. in Biology, Senior Research Fellow, Department of Virological and Ultrastructural Research, Federal Center for Toxicological, Radiation and Biological Safety; st. Scientific town-2, Kazan, 420075; phone: 8-987-4214127; e-mail: tanirtashir@mail.ru.

Khafizova Alsu M., Junior Research Fellow, Department of Virological and Ultrastructural Research, Federal Center for Toxicological, Radiation and Biological Safety; st. Scientific town-2, Kazan, 420075; phone: 9-927-4294978; e-mail: alsukhafizova@yandex.ru.

Responsible for correspondence with the editorial board: Mukhammadiev Rishat S., Ph.D. in Biology, Senior Research Fellow, Department of Virological and Ultrastructural Research, Federal Center for Toxicological, Radiation and Biological Safety; st. Scientific town-2, Kazan, 420075; phone: 8-939-3728789; e-mail: tashir9891@mail.ru.

Authors’ Contribution:

Mukhammadiev Rish.S.: conceptualization, supervision, writing - editing.

Karimullina I.G.: work administration, writing - original draft preparation

Yarullin A.I.: data curation, resources.

Sorokina D.A.: investigation, validation.

Khaertynov K.S.: methodology, investigation.

Galeeva A.G.: formal analysis.

Mukhammadiev Rin.S.: validation, visualization.

Khafizova A.M.: investigation.

Conflict of Interest Statement: the authors declare no conflict of interest.