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Preparation and Biochemical Characteristics of a New IgG-Type Monoclonal Antibody against K Subgroup Avian Leukosis Virus

[Image: see text] This study focused on preparing a new IgG-type monoclonal antibody (MAb) against subgroup K avian leukosis virus (ALV-K) and identifying its biochemical characteristics. A specific gene fragment of ALV-K was amplified by polymerase chain reaction and expressed in E. coli. The purif...

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Autores principales: Zhang, Xiaochen, Li, Hongmei, Wang, Chengcheng, Du, Yixuan, Li, Yuying, Zhang, Liwei, Huang, Mengjie, Qiu, Jianhua, Guo, Huijun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2022
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9835519/
https://www.ncbi.nlm.nih.gov/pubmed/36643488
http://dx.doi.org/10.1021/acsomega.2c06375
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author Zhang, Xiaochen
Li, Hongmei
Wang, Chengcheng
Du, Yixuan
Li, Yuying
Zhang, Liwei
Huang, Mengjie
Qiu, Jianhua
Guo, Huijun
author_facet Zhang, Xiaochen
Li, Hongmei
Wang, Chengcheng
Du, Yixuan
Li, Yuying
Zhang, Liwei
Huang, Mengjie
Qiu, Jianhua
Guo, Huijun
author_sort Zhang, Xiaochen
collection PubMed
description [Image: see text] This study focused on preparing a new IgG-type monoclonal antibody (MAb) against subgroup K avian leukosis virus (ALV-K) and identifying its biochemical characteristics. A specific gene fragment of ALV-K was amplified by polymerase chain reaction and expressed in E. coli. The purified expressed products were inoculated into BALB/c mice to prepare antibody-secreting spleen lymphocytes, and hybridoma cells were obtained after cell fusion of spleen lymphocytes and myeloma cells. A new hybridoma cell line named 30B9, which stably secreted IgG2b-antibody against ALV-K, was screened and contained 98 chromosomes. The MAb secreted by the 30B9 cells could recognize the ALV-K strain but not the ALV-A/B/J strains in an indirect immunofluorescence assay. Seventeen overlapping truncated ALV-K gp85 protein fragments were expressed, and eight peptides were artificially synthesized to analyze the MAb’s antigen epitope by Western blot or enzyme-linked immunosorbent assay, and the results showed that the linear epitope was located on the (217–)RRNYT(–221) of ALV-K gp85 protein. A bioinformatics analysis showed that the epitope has a high antigenicity index, hydrophilicity, and surface accessibility and forms a unique linear spatial structure. Its five amino acids are highly conserved in all published ALV-K strains but are very low in ALV-A/B/J/C/D/E strains. This study provides a new biomaterial for developing specific detection methods against ALV-K.
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spelling pubmed-98355192023-01-13 Preparation and Biochemical Characteristics of a New IgG-Type Monoclonal Antibody against K Subgroup Avian Leukosis Virus Zhang, Xiaochen Li, Hongmei Wang, Chengcheng Du, Yixuan Li, Yuying Zhang, Liwei Huang, Mengjie Qiu, Jianhua Guo, Huijun ACS Omega [Image: see text] This study focused on preparing a new IgG-type monoclonal antibody (MAb) against subgroup K avian leukosis virus (ALV-K) and identifying its biochemical characteristics. A specific gene fragment of ALV-K was amplified by polymerase chain reaction and expressed in E. coli. The purified expressed products were inoculated into BALB/c mice to prepare antibody-secreting spleen lymphocytes, and hybridoma cells were obtained after cell fusion of spleen lymphocytes and myeloma cells. A new hybridoma cell line named 30B9, which stably secreted IgG2b-antibody against ALV-K, was screened and contained 98 chromosomes. The MAb secreted by the 30B9 cells could recognize the ALV-K strain but not the ALV-A/B/J strains in an indirect immunofluorescence assay. Seventeen overlapping truncated ALV-K gp85 protein fragments were expressed, and eight peptides were artificially synthesized to analyze the MAb’s antigen epitope by Western blot or enzyme-linked immunosorbent assay, and the results showed that the linear epitope was located on the (217–)RRNYT(–221) of ALV-K gp85 protein. A bioinformatics analysis showed that the epitope has a high antigenicity index, hydrophilicity, and surface accessibility and forms a unique linear spatial structure. Its five amino acids are highly conserved in all published ALV-K strains but are very low in ALV-A/B/J/C/D/E strains. This study provides a new biomaterial for developing specific detection methods against ALV-K. American Chemical Society 2022-12-22 /pmc/articles/PMC9835519/ /pubmed/36643488 http://dx.doi.org/10.1021/acsomega.2c06375 Text en © 2022 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by-nc-nd/4.0/Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Zhang, Xiaochen
Li, Hongmei
Wang, Chengcheng
Du, Yixuan
Li, Yuying
Zhang, Liwei
Huang, Mengjie
Qiu, Jianhua
Guo, Huijun
Preparation and Biochemical Characteristics of a New IgG-Type Monoclonal Antibody against K Subgroup Avian Leukosis Virus
title Preparation and Biochemical Characteristics of a New IgG-Type Monoclonal Antibody against K Subgroup Avian Leukosis Virus
title_full Preparation and Biochemical Characteristics of a New IgG-Type Monoclonal Antibody against K Subgroup Avian Leukosis Virus
title_fullStr Preparation and Biochemical Characteristics of a New IgG-Type Monoclonal Antibody against K Subgroup Avian Leukosis Virus
title_full_unstemmed Preparation and Biochemical Characteristics of a New IgG-Type Monoclonal Antibody against K Subgroup Avian Leukosis Virus
title_short Preparation and Biochemical Characteristics of a New IgG-Type Monoclonal Antibody against K Subgroup Avian Leukosis Virus
title_sort preparation and biochemical characteristics of a new igg-type monoclonal antibody against k subgroup avian leukosis virus
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9835519/
https://www.ncbi.nlm.nih.gov/pubmed/36643488
http://dx.doi.org/10.1021/acsomega.2c06375
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