Identification of Two Novel Linear Neutralizing Epitopes within the Hexon Protein of Canine Adenovirus Using Monoclonal Antibodies
Canine adenovirus (CAdV) has a high prevalence in canine populations. High affinity neutralizing antibodies against conserved epitopes can provide protective immunity against CAdV and protect against future outbreaks. In this study, we identified two CAdV-2-specific neutralizing monoclonal antibodie...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-02-01
|
| Series: | Vaccines |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-393X/9/2/135 |
| _version_ | 1797412631852613632 |
|---|---|
| author | Shujie Wang Chunsheng Wang Xiao Ren Wenjiao Xue Haijuan He Yanzhu Zhu Hongfeng Wang Gang Wang Xuehui Cai |
| author_facet | Shujie Wang Chunsheng Wang Xiao Ren Wenjiao Xue Haijuan He Yanzhu Zhu Hongfeng Wang Gang Wang Xuehui Cai |
| author_sort | Shujie Wang |
| collection | DOAJ |
| description | Canine adenovirus (CAdV) has a high prevalence in canine populations. High affinity neutralizing antibodies against conserved epitopes can provide protective immunity against CAdV and protect against future outbreaks. In this study, we identified two CAdV-2-specific neutralizing monoclonal antibodies (mAbs), 2C1 and 7D7, which recognized two linear-dependent epitopes. MAb 2C1 potently neutralized CAdV-2 with a 50% neutralization titer (NT50) of 4096, and mAb 7D7 partially neutralized CAdV-2 with a 50% NT50 of 64. Immunoprecipitation, Western blot and protein spectral analysis indicated that both neutralizing mAbs recognized the hexon protein (Hex) of CAdV-2. Through a 12-mer random peptide phage display and synthetic peptides analysis, we finely mapped the neutralizing epitopes to two 10-amino acid (aa) peptides within the CAdV Hex: <sup>634</sup>RIKQRETPAL<sup>643</sup> located on the surface region; and <sup>736</sup>PESYKDRMYS<sup>745</sup> located in the inner region of the expected 3D structure of trimeric Hex. Importantly, the two epitopes are highly conserved among all CAdV isolates by sequence alignment analysis. Thus, these results provide insights into the interaction between virus and mAbs at the aa level and may have potential applications in the development of novel therapeutic or epitope-based vaccines, antibody therapeutics and a diagnostic method suitable for the rapid detection of all CAdVs. |
| first_indexed | 2024-03-09T05:05:55Z |
| format | Article |
| id | doaj.art-51383a4203824531813c47d9facbc429 |
| institution | Directory Open Access Journal |
| issn | 2076-393X |
| language | English |
| last_indexed | 2024-03-09T05:05:55Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj.art-51383a4203824531813c47d9facbc4292023-12-03T12:55:07ZengMDPI AGVaccines2076-393X2021-02-019213510.3390/vaccines9020135Identification of Two Novel Linear Neutralizing Epitopes within the Hexon Protein of Canine Adenovirus Using Monoclonal AntibodiesShujie Wang0Chunsheng Wang1Xiao Ren2Wenjiao Xue3Haijuan He4Yanzhu Zhu5Hongfeng Wang6Gang Wang7Xuehui Cai8State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, ChinaCollege of Life Science, Northeast Forestry University, Harbin 150040, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, ChinaKey Laboratory of Special Animal Epidemic Disease, Ministry of Agriculture, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, ChinaCanine adenovirus (CAdV) has a high prevalence in canine populations. High affinity neutralizing antibodies against conserved epitopes can provide protective immunity against CAdV and protect against future outbreaks. In this study, we identified two CAdV-2-specific neutralizing monoclonal antibodies (mAbs), 2C1 and 7D7, which recognized two linear-dependent epitopes. MAb 2C1 potently neutralized CAdV-2 with a 50% neutralization titer (NT50) of 4096, and mAb 7D7 partially neutralized CAdV-2 with a 50% NT50 of 64. Immunoprecipitation, Western blot and protein spectral analysis indicated that both neutralizing mAbs recognized the hexon protein (Hex) of CAdV-2. Through a 12-mer random peptide phage display and synthetic peptides analysis, we finely mapped the neutralizing epitopes to two 10-amino acid (aa) peptides within the CAdV Hex: <sup>634</sup>RIKQRETPAL<sup>643</sup> located on the surface region; and <sup>736</sup>PESYKDRMYS<sup>745</sup> located in the inner region of the expected 3D structure of trimeric Hex. Importantly, the two epitopes are highly conserved among all CAdV isolates by sequence alignment analysis. Thus, these results provide insights into the interaction between virus and mAbs at the aa level and may have potential applications in the development of novel therapeutic or epitope-based vaccines, antibody therapeutics and a diagnostic method suitable for the rapid detection of all CAdVs.https://www.mdpi.com/2076-393X/9/2/135canine adenovirusidentificationhexon proteinmonoclonal antibodiesnovel epitopes |
| spellingShingle | Shujie Wang Chunsheng Wang Xiao Ren Wenjiao Xue Haijuan He Yanzhu Zhu Hongfeng Wang Gang Wang Xuehui Cai Identification of Two Novel Linear Neutralizing Epitopes within the Hexon Protein of Canine Adenovirus Using Monoclonal Antibodies Vaccines canine adenovirus identification hexon protein monoclonal antibodies novel epitopes |
| title | Identification of Two Novel Linear Neutralizing Epitopes within the Hexon Protein of Canine Adenovirus Using Monoclonal Antibodies |
| title_full | Identification of Two Novel Linear Neutralizing Epitopes within the Hexon Protein of Canine Adenovirus Using Monoclonal Antibodies |
| title_fullStr | Identification of Two Novel Linear Neutralizing Epitopes within the Hexon Protein of Canine Adenovirus Using Monoclonal Antibodies |
| title_full_unstemmed | Identification of Two Novel Linear Neutralizing Epitopes within the Hexon Protein of Canine Adenovirus Using Monoclonal Antibodies |
| title_short | Identification of Two Novel Linear Neutralizing Epitopes within the Hexon Protein of Canine Adenovirus Using Monoclonal Antibodies |
| title_sort | identification of two novel linear neutralizing epitopes within the hexon protein of canine adenovirus using monoclonal antibodies |
| topic | canine adenovirus identification hexon protein monoclonal antibodies novel epitopes |
| url | https://www.mdpi.com/2076-393X/9/2/135 |
| work_keys_str_mv | AT shujiewang identificationoftwonovellinearneutralizingepitopeswithinthehexonproteinofcanineadenovirususingmonoclonalantibodies AT chunshengwang identificationoftwonovellinearneutralizingepitopeswithinthehexonproteinofcanineadenovirususingmonoclonalantibodies AT xiaoren identificationoftwonovellinearneutralizingepitopeswithinthehexonproteinofcanineadenovirususingmonoclonalantibodies AT wenjiaoxue identificationoftwonovellinearneutralizingepitopeswithinthehexonproteinofcanineadenovirususingmonoclonalantibodies AT haijuanhe identificationoftwonovellinearneutralizingepitopeswithinthehexonproteinofcanineadenovirususingmonoclonalantibodies AT yanzhuzhu identificationoftwonovellinearneutralizingepitopeswithinthehexonproteinofcanineadenovirususingmonoclonalantibodies AT hongfengwang identificationoftwonovellinearneutralizingepitopeswithinthehexonproteinofcanineadenovirususingmonoclonalantibodies AT gangwang identificationoftwonovellinearneutralizingepitopeswithinthehexonproteinofcanineadenovirususingmonoclonalantibodies AT xuehuicai identificationoftwonovellinearneutralizingepitopeswithinthehexonproteinofcanineadenovirususingmonoclonalantibodies |