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Article

Expression and Immunological Characterization of African Swine Fever Virus EP153R Protein for Serodiagnosis and Its Delivery via a Recombinant PRRSV Live Vector

by
Meng Luo
1,2,†,
Wenna Shuai
1,†,
Ziqiang Guo
1,
Jiale Li
1,
Liwei Li
1,
Yanjun Zhou
1,
Yifeng Jiang
1,
Wu Tong
1,
Yifan Zeng
1,
Jinbin Wang
1,
Li Zhao
3,4 and
Fei Gao
1,5,*
1
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China
2
College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China
3
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
4
Ruipu Biological Co., Ltd., Tianjin 300308, China
5
Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonose, Yangzhou University, Yangzhou 225009, China
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Vaccines 2025, 13(11), 1110; https://doi.org/10.3390/vaccines13111110
Submission received: 25 September 2025 / Revised: 23 October 2025 / Accepted: 27 October 2025 / Published: 29 October 2025
(This article belongs to the Special Issue The Immunotherapy Against Swine Disease)
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Abstract

Background/Objectives: African Swine Fever (ASF), caused by the African Swine Fever Virus (ASFV), is a highly contagious and lethal disease in pigs, for which no recognized safe and effective vaccine is currently available. The ASFV EP153R gene, expressed during both early and late infection stages, exhibits strong protective potential. Utilizing advances in genetic engineering, recombinant PRRSV vector vaccines carrying ASFV exogenous genes were constructed. This study aims to prepare pEP153R-based polyclonal antibodies and an iELISA detection method using the constructed rPRRSV-EP153R as a specific target to verify the iELISA’s specificity and effectiveness. Methods: A prokaryotic plasmid, pCold-TF-EP153R, was constructed to express protein in BL21 (DE3). The purified soluble protein (2 mg/mL) was used to generate a murine polyclonal antibody and establish an indirect ELISA. The EP153R gene was inserted between ORF1b and ORF2a of PRRSV via reverse genetics, yielding recombinant rPRRSV-EP153R. Its biological properties were assessed in vitro and in vivo. Results: The pEP153R was specifically detected by both anti-His antibody and generated polyclonal antibodies. An established iELISA showed high specificity, sensitivity, and 98.18% accuracy. The antibodies specifically recognized pEP153R expressed in recombinant virus and eukaryotic systems. Additionally, the recombinant virus stably maintained EP153R without changes in virological characteristics relative to vHuN4-F112. In vaccinated piglets, the rPRRSV-EP153R induced a specific, consistent, and detectable immune response. Conclusions: The established iELISA, characterized by high specificity, sensitivity, and accuracy, furnishes reliable technical support for the serological diagnosis of ASFV. Meanwhile, the recombinant virus rPRRSV-EP153R demonstrates potential as a novel live vectored vaccine candidate, with the capability to induce specific immunity against both ASFV and PRRSV.
Keywords: African swine fever virus; pEP153R; recombinant protein expression; immunogenicity evaluation; polyclonal antibody production; serodiagnosis; indirect ELISA; PRRSV-based viral vector; reverse genetics; viral live vector African swine fever virus; pEP153R; recombinant protein expression; immunogenicity evaluation; polyclonal antibody production; serodiagnosis; indirect ELISA; PRRSV-based viral vector; reverse genetics; viral live vector

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MDPI and ACS Style

Luo, M.; Shuai, W.; Guo, Z.; Li, J.; Li, L.; Zhou, Y.; Jiang, Y.; Tong, W.; Zeng, Y.; Wang, J.; et al. Expression and Immunological Characterization of African Swine Fever Virus EP153R Protein for Serodiagnosis and Its Delivery via a Recombinant PRRSV Live Vector. Vaccines 2025, 13, 1110. https://doi.org/10.3390/vaccines13111110

AMA Style

Luo M, Shuai W, Guo Z, Li J, Li L, Zhou Y, Jiang Y, Tong W, Zeng Y, Wang J, et al. Expression and Immunological Characterization of African Swine Fever Virus EP153R Protein for Serodiagnosis and Its Delivery via a Recombinant PRRSV Live Vector. Vaccines. 2025; 13(11):1110. https://doi.org/10.3390/vaccines13111110

Chicago/Turabian Style

Luo, Meng, Wenna Shuai, Ziqiang Guo, Jiale Li, Liwei Li, Yanjun Zhou, Yifeng Jiang, Wu Tong, Yifan Zeng, Jinbin Wang, and et al. 2025. "Expression and Immunological Characterization of African Swine Fever Virus EP153R Protein for Serodiagnosis and Its Delivery via a Recombinant PRRSV Live Vector" Vaccines 13, no. 11: 1110. https://doi.org/10.3390/vaccines13111110

APA Style

Luo, M., Shuai, W., Guo, Z., Li, J., Li, L., Zhou, Y., Jiang, Y., Tong, W., Zeng, Y., Wang, J., Zhao, L., & Gao, F. (2025). Expression and Immunological Characterization of African Swine Fever Virus EP153R Protein for Serodiagnosis and Its Delivery via a Recombinant PRRSV Live Vector. Vaccines, 13(11), 1110. https://doi.org/10.3390/vaccines13111110

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