Optimization of viral antigen purification for the production of immunological agents used for treatment and prevention of young livestock infections

Document Type : Research Paper

Authors

1 Scientific Production Technical Center "Zhalyn", Almaty, Kazakhstan

2 Neurox Technology Ltd., Türkiye

3 Institute of Genetics and Physiology, Almaty, Kazakhstan

10.22124/cjes.2026.10053

Abstract

Gastrointestinal diseases in neonatal calves cause significant economic losses to the agricultural sector worldwide. Commonly identified causative agents include viral pathogens such as bovine viral diarrhea virus (BVDV), bovine rotavirus group A (BRVA), bovine coronavirus (BCoV), bovine alpha-herpesvirus type 1 (BoAHV-1), and bovine parainfluenza virus type 3 (BPI3V). Clinical manifestations of the infections include diarrhea, dehydration, weight loss, reduced milk production, and, in severe cases, fatal outcomes, primarily among calves under one month of life, with mortality rates reaching 50%. The production of effective hyperimmune sera and immunological drugs requires high-quality viral antigens with enhanced immunogenic properties, which determines the need to develop optimized technologies for virus purification and concentration. This study was conducted in 2025 at the “Biogenic Preparations” laboratory of the “Zhalyn” Scientific-Production Technological Center in Almaty, Kazakhstan, to evaluate and optimize various methods for purifying viral antigens used in the production of immunological preparations. Five major viral pathogens of cattle were cultivated in MDBK cell cultures and treated with four different concentration methods: precipitation with polyethylene glycol (PEG) of various molecular weights, ammonium sulfate precipitation, ultrafiltration, and high-speed ultracentrifugation.
The experimental results demonstrated all tested methods increased the concentration of viral particles and enhanced the infectious activity of the suspension, in varying degrees. Precipitation with PEG molecular weight 6,000 at a concentration of 7.5% yielded optimal results, providing a significant increase in infectious activity of 2 log units. Precipitation with ammonium sulfate led to a moderate increase in concentration with an enhancement of viral activity by approximately 1 log unit. Ultrafiltration demonstrated exceptional efficacy, achieving concentration factors ranging from 10 to 50 times with maximum activity values comparable to PEG purification, while providing the advantages of purification through the removal of minor impurities. Ultracentrifugation, while demonstrating a less significant increase in titer (by 0.5–0.7 log units), provided superior purification quality, which is critical for high-quality therapeutic products. Statistical analysis using ANOVA and Tukey’s multiple comparison tests confirmed the significance of the differences observed (p < 0.05). These findings demonstrate that ultrafiltration and PEG precipitation represent the most effective methods for increasing viral titer with minimal loss of biological activity, making them ideal for large-scale production of diagnostic and therapeutic products. Ultracentrifugation, despite lower concentration rates, provides levels of purity essential for research and specialized therapeutic applications. The study demonstrates that an integrated strategy combining multiple concentration methods optimizes both the quality and activity of viral preparations. The study’s findings present opportunities for optimizing biotechnology production and veterinary pharmaceuticals by standardizing protocols for the concentration of viral antigens used to prevent and treat gastrointestinal diseases in cattle.

Keywords

References

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Caspian Journal of Environmental Sciences
Volume 24, Issue 4
October 2026
Pages 1209-1217

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