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Preparation of a novel monoclonal antibody against Avian leukosis virus subgroup J Gp85 protein and identification of its epitope

Avian leukosis virus subgroup J (ALV-J) is an avian oncogenic retrovirus that has caused huge economic losses in the poultry industry due to its great pathogenicity and transmission ability. However, the continuous emergence of new strains would bring challenges to diagnosis and control of ALV-J. .T...

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Autores principales: Wang, Houkun, Chen, Xueyang, Zhu, Lilin, Fang, Xiaowei, Gao, Keli, Fang, Chun, Liu, Jing, Gu, Yufang, Liang, Xiongyan, Yang, Yuying
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8192869/
https://www.ncbi.nlm.nih.gov/pubmed/34116348
http://dx.doi.org/10.1016/j.psj.2021.101108
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author Wang, Houkun
Chen, Xueyang
Zhu, Lilin
Fang, Xiaowei
Gao, Keli
Fang, Chun
Liu, Jing
Gu, Yufang
Liang, Xiongyan
Yang, Yuying
author_facet Wang, Houkun
Chen, Xueyang
Zhu, Lilin
Fang, Xiaowei
Gao, Keli
Fang, Chun
Liu, Jing
Gu, Yufang
Liang, Xiongyan
Yang, Yuying
author_sort Wang, Houkun
collection PubMed
description Avian leukosis virus subgroup J (ALV-J) is an avian oncogenic retrovirus that has caused huge economic losses in the poultry industry due to its great pathogenicity and transmission ability. However, the continuous emergence of new strains would bring challenges to diagnosis and control of ALV-J. .This study focuses on preparing the monoclonal antibody (MAb) against ALV-J Gp85 and identifying its epitope. The truncated ALV-J gp85 gene fragment was amplified and then cloned into expression vectors. Purified GST-Gp85 was used to immune mice and His-Gp85 was used to screen MAb. Finally, a hybridoma cell line named J16 that produced specific MAb against the ALV-J. Immunofluorescence assay showed that MAb J16 specifically recognized ALV-J rather than ALV-A or ALV-K infected DF-1 cells. To identify the epitope recognized by MAb J16, fourteen partially overlapping ALV-J Gp85 fragments were prepared and tested by Western blot. The results indicated that peptide 150-LIRPYVNQ-157 was the minimal epitope of ALV-J Gp85 recognized by MAb J16. Alignment analysis of Gp85 from different ALV subgroups showed that the epitope keep high conservation among 36 ALV-J strains, but significant different from that of ALV subgroup A, B, C, D, E and K. Overall, we prepared a MAb specific against ALV-J and identified peptide 150-LIRPYVNQ-157 as a novel specific epitope of ALV-J Gp85, which may assist in laying the foundation for specific ALV-J detection methods.
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spelling pubmed-81928692021-06-17 Preparation of a novel monoclonal antibody against Avian leukosis virus subgroup J Gp85 protein and identification of its epitope Wang, Houkun Chen, Xueyang Zhu, Lilin Fang, Xiaowei Gao, Keli Fang, Chun Liu, Jing Gu, Yufang Liang, Xiongyan Yang, Yuying Poult Sci IMMUNOLOGY, HEALTH AND DISEASE Avian leukosis virus subgroup J (ALV-J) is an avian oncogenic retrovirus that has caused huge economic losses in the poultry industry due to its great pathogenicity and transmission ability. However, the continuous emergence of new strains would bring challenges to diagnosis and control of ALV-J. .This study focuses on preparing the monoclonal antibody (MAb) against ALV-J Gp85 and identifying its epitope. The truncated ALV-J gp85 gene fragment was amplified and then cloned into expression vectors. Purified GST-Gp85 was used to immune mice and His-Gp85 was used to screen MAb. Finally, a hybridoma cell line named J16 that produced specific MAb against the ALV-J. Immunofluorescence assay showed that MAb J16 specifically recognized ALV-J rather than ALV-A or ALV-K infected DF-1 cells. To identify the epitope recognized by MAb J16, fourteen partially overlapping ALV-J Gp85 fragments were prepared and tested by Western blot. The results indicated that peptide 150-LIRPYVNQ-157 was the minimal epitope of ALV-J Gp85 recognized by MAb J16. Alignment analysis of Gp85 from different ALV subgroups showed that the epitope keep high conservation among 36 ALV-J strains, but significant different from that of ALV subgroup A, B, C, D, E and K. Overall, we prepared a MAb specific against ALV-J and identified peptide 150-LIRPYVNQ-157 as a novel specific epitope of ALV-J Gp85, which may assist in laying the foundation for specific ALV-J detection methods. Elsevier 2021-03-12 /pmc/articles/PMC8192869/ /pubmed/34116348 http://dx.doi.org/10.1016/j.psj.2021.101108 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle IMMUNOLOGY, HEALTH AND DISEASE
Wang, Houkun
Chen, Xueyang
Zhu, Lilin
Fang, Xiaowei
Gao, Keli
Fang, Chun
Liu, Jing
Gu, Yufang
Liang, Xiongyan
Yang, Yuying
Preparation of a novel monoclonal antibody against Avian leukosis virus subgroup J Gp85 protein and identification of its epitope
title Preparation of a novel monoclonal antibody against Avian leukosis virus subgroup J Gp85 protein and identification of its epitope
title_full Preparation of a novel monoclonal antibody against Avian leukosis virus subgroup J Gp85 protein and identification of its epitope
title_fullStr Preparation of a novel monoclonal antibody against Avian leukosis virus subgroup J Gp85 protein and identification of its epitope
title_full_unstemmed Preparation of a novel monoclonal antibody against Avian leukosis virus subgroup J Gp85 protein and identification of its epitope
title_short Preparation of a novel monoclonal antibody against Avian leukosis virus subgroup J Gp85 protein and identification of its epitope
title_sort preparation of a novel monoclonal antibody against avian leukosis virus subgroup j gp85 protein and identification of its epitope
topic IMMUNOLOGY, HEALTH AND DISEASE
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8192869/
https://www.ncbi.nlm.nih.gov/pubmed/34116348
http://dx.doi.org/10.1016/j.psj.2021.101108
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