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Isolation and Characterization of Monoclonal Antibodies Against a Virion Core Protein of Orf Virus Strain NA1/11 As Potential Diagnostic Tool for Orf Viruses.

Abstract

Orf is caused by the orf virus (ORFV) and is a non-systemic, widespread disease afflicting sheep, goats, wild ruminants, and humans. Recent outbreaks in sheep and goats in Jilin and other northern Chinese provinces raise concerns about orf control in China. Thirty-five hybridoma clones were constructed from splenocytes of BALB/c mice immunized with natural orf virus protein. These hybridomas were used to produce antibodies targeting ORFV proteins. Immunological characterization of these monoclonal antibodies (MAb) showed that the 5F2D8 hybridoma line produced MAb that can recognize the 100, 70, and 20 kDa bands from total viral lysate. This hybridoma was further characterized by immunoprecipitation and peptide sequencing. The results indicate that 5F2D8 specifically recognizes orf virus encoded protein ORFV086, a late expression virion core protein that plays important roles in progeny virus particle assembly, morphogenesis, and maturity. Further experiments demonstrate that this MAb did not react with other viral proteins of ORFV orthopoxviruses, but reacted strongly to different field isolates of orf viruses from China. Additionally, this anti-ORFV086 MAb possesses ORFV neutralizing capability. Sequence alignments and phylogenetic analysis determined that ORFV086 of NA1/11, clustered together with NZ2 and IA82, is highly conserved and has structural similarities with the Vaccinia virus core protein P4a. As such, this MAb has great potential as a diagnostic tool for orf viruses, in the further exploration of orf pathogenesis, and in disease control and prevention.

Authors+Show Affiliations

Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, People's Republic of China .Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, People's Republic of China .Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, People's Republic of China .Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, People's Republic of China .Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, People's Republic of China .Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, People's Republic of China .Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, People's Republic of China .Institute of Antibody Engineering, School of Biotechnology, Southern Medical University , Guangzhou, People's Republic of China .

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

26301926

Citation

Wang, Xiaoping, et al. "Isolation and Characterization of Monoclonal Antibodies Against a Virion Core Protein of Orf Virus Strain NA1/11 as Potential Diagnostic Tool for Orf Viruses." Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, vol. 34, no. 4, 2015, pp. 233-45.
Wang X, Zhang J, Hao W, et al. Isolation and Characterization of Monoclonal Antibodies Against a Virion Core Protein of Orf Virus Strain NA1/11 As Potential Diagnostic Tool for Orf Viruses. Monoclon Antib Immunodiagn Immunother. 2015;34(4):233-45.
Wang, X., Zhang, J., Hao, W., Peng, Y., Li, H., Li, W., ... Luo, S. (2015). Isolation and Characterization of Monoclonal Antibodies Against a Virion Core Protein of Orf Virus Strain NA1/11 As Potential Diagnostic Tool for Orf Viruses. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, 34(4), pp. 233-45. doi:10.1089/mab.2014.0101.
Wang X, et al. Isolation and Characterization of Monoclonal Antibodies Against a Virion Core Protein of Orf Virus Strain NA1/11 as Potential Diagnostic Tool for Orf Viruses. Monoclon Antib Immunodiagn Immunother. 2015;34(4):233-45. PubMed PMID: 26301926.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Isolation and Characterization of Monoclonal Antibodies Against a Virion Core Protein of Orf Virus Strain NA1/11 As Potential Diagnostic Tool for Orf Viruses. AU - Wang,Xiaoping, AU - Zhang,Jiafeng, AU - Hao,Wenbo, AU - Peng,Yongzheng, AU - Li,Hong, AU - Li,Wei, AU - Li,Ming, AU - Luo,Shuhong, PY - 2015/8/25/entrez PY - 2015/8/25/pubmed PY - 2016/5/19/medline SP - 233 EP - 45 JF - Monoclonal antibodies in immunodiagnosis and immunotherapy JO - Monoclon Antib Immunodiagn Immunother VL - 34 IS - 4 N2 - Orf is caused by the orf virus (ORFV) and is a non-systemic, widespread disease afflicting sheep, goats, wild ruminants, and humans. Recent outbreaks in sheep and goats in Jilin and other northern Chinese provinces raise concerns about orf control in China. Thirty-five hybridoma clones were constructed from splenocytes of BALB/c mice immunized with natural orf virus protein. These hybridomas were used to produce antibodies targeting ORFV proteins. Immunological characterization of these monoclonal antibodies (MAb) showed that the 5F2D8 hybridoma line produced MAb that can recognize the 100, 70, and 20 kDa bands from total viral lysate. This hybridoma was further characterized by immunoprecipitation and peptide sequencing. The results indicate that 5F2D8 specifically recognizes orf virus encoded protein ORFV086, a late expression virion core protein that plays important roles in progeny virus particle assembly, morphogenesis, and maturity. Further experiments demonstrate that this MAb did not react with other viral proteins of ORFV orthopoxviruses, but reacted strongly to different field isolates of orf viruses from China. Additionally, this anti-ORFV086 MAb possesses ORFV neutralizing capability. Sequence alignments and phylogenetic analysis determined that ORFV086 of NA1/11, clustered together with NZ2 and IA82, is highly conserved and has structural similarities with the Vaccinia virus core protein P4a. As such, this MAb has great potential as a diagnostic tool for orf viruses, in the further exploration of orf pathogenesis, and in disease control and prevention. SN - 2167-9436 UR - https://www.unboundmedicine.com/medline/citation/26301926/Isolation_and_Characterization_of_Monoclonal_Antibodies_Against_a_Virion_Core_Protein_of_Orf_Virus_Strain_NA1/11_As_Potential_Diagnostic_Tool_for_Orf_Viruses_ L2 - https://www.liebertpub.com/doi/full/10.1089/mab.2014.0101?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -