A highly efficient indirect ELISA and monoclonal antibody established against African swine fever virus pK205R
African swine fever (ASF) is a contagious infectious disease with high lethality which continuously threatens the global pig industry causing huge economic losses. Currently, there are no commercially available vaccines or antiviral drugs that can effectively control ASF. The pathogen of ASF, ASF vi...
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| Published in | Frontiers in immunology Vol. 13; p. 1103166 |
|---|---|
| Main Authors | , , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Switzerland
Frontiers Media S.A
09.01.2023
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1664-3224 1664-3224 |
| DOI | 10.3389/fimmu.2022.1103166 |
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| Abstract | African swine fever (ASF) is a contagious infectious disease with high lethality which continuously threatens the global pig industry causing huge economic losses. Currently, there are no commercially available vaccines or antiviral drugs that can effectively control ASF. The pathogen of ASF, ASF virus (ASFV) is a double-stranded DNA virus with a genome ranging from 170 to 193 kb and 151 to 167 open reading frames in various strains, which encodes 150–200 proteins. An effective method of monitoring ASFV antibodies, and specific antibodies against ASFV to promote the development of prevention techniques are urgently needed. In the present study, pK205R of ASFV was successfully expressed in mammalian cells using a suspension culture system. An indirect enzyme-linked immunosorbent assay (ELISA) based on the purified pK205R was established and optimized. The monoclonal antibody (mAb) against pK205R recognized a conservative linear epitope (
2
VEPREQFFQDLLSAV
16
) and exhibited specific reactivity, which was conducive to the identification of the recombinant porcine reproductive and respiratory syndrome virus (PRRSV) expressing pK205R. The ELISA method efficiently detected clinical ASFV infection and revealed good application prospects in monitoring the antibody level
in vivo
for recombinant PRRSV live vector virus expressing the ASFV antigen protein. The determination of the conserved linear epitope of pK205R would contribute to further research on the structural biology and function of pK205R. The indirect ELISA method and mAb against ASFV pK205R revealed efficient detection and promising application prospects, making them ideal for epidemiological surveillance and vaccine research on ASF. |
|---|---|
| AbstractList | African swine fever (ASF) is a contagious infectious disease with high lethality which continuously threatens the global pig industry causing huge economic losses. Currently, there are no commercially available vaccines or antiviral drugs that can effectively control ASF. The pathogen of ASF, ASF virus (ASFV) is a double-stranded DNA virus with a genome ranging from 170 to 193 kb and 151 to 167 open reading frames in various strains, which encodes 150-200 proteins. An effective method of monitoring ASFV antibodies, and specific antibodies against ASFV to promote the development of prevention techniques are urgently needed. In the present study, pK205R of ASFV was successfully expressed in mammalian cells using a suspension culture system. An indirect enzyme-linked immunosorbent assay (ELISA) based on the purified pK205R was established and optimized. The monoclonal antibody (mAb) against pK205R recognized a conservative linear epitope (
VEPREQFFQDLLSAV
) and exhibited specific reactivity, which was conducive to the identification of the recombinant porcine reproductive and respiratory syndrome virus (PRRSV) expressing pK205R. The ELISA method efficiently detected clinical ASFV infection and revealed good application prospects in monitoring the antibody level
for recombinant PRRSV live vector virus expressing the ASFV antigen protein. The determination of the conserved linear epitope of pK205R would contribute to further research on the structural biology and function of pK205R. The indirect ELISA method and mAb against ASFV pK205R revealed efficient detection and promising application prospects, making them ideal for epidemiological surveillance and vaccine research on ASF. African swine fever (ASF) is a contagious infectious disease with high lethality which continuously threatens the global pig industry causing huge economic losses. Currently, there are no commercially available vaccines or antiviral drugs that can effectively control ASF. The pathogen of ASF, ASF virus (ASFV) is a double-stranded DNA virus with a genome ranging from 170 to 193 kb and 151 to 167 open reading frames in various strains, which encodes 150–200 proteins. An effective method of monitoring ASFV antibodies, and specific antibodies against ASFV to promote the development of prevention techniques are urgently needed. In the present study, pK205R of ASFV was successfully expressed in mammalian cells using a suspension culture system. An indirect enzyme-linked immunosorbent assay (ELISA) based on the purified pK205R was established and optimized. The monoclonal antibody (mAb) against pK205R recognized a conservative linear epitope ( 2 VEPREQFFQDLLSAV 16 ) and exhibited specific reactivity, which was conducive to the identification of the recombinant porcine reproductive and respiratory syndrome virus (PRRSV) expressing pK205R. The ELISA method efficiently detected clinical ASFV infection and revealed good application prospects in monitoring the antibody level in vivo for recombinant PRRSV live vector virus expressing the ASFV antigen protein. The determination of the conserved linear epitope of pK205R would contribute to further research on the structural biology and function of pK205R. The indirect ELISA method and mAb against ASFV pK205R revealed efficient detection and promising application prospects, making them ideal for epidemiological surveillance and vaccine research on ASF. African swine fever (ASF) is a contagious infectious disease with high lethality which continuously threatens the global pig industry causing huge economic losses. Currently, there are no commercially available vaccines or antiviral drugs that can effectively control ASF. The pathogen of ASF, ASF virus (ASFV) is a double-stranded DNA virus with a genome ranging from 170 to 193 kb and 151 to 167 open reading frames in various strains, which encodes 150-200 proteins. An effective method of monitoring ASFV antibodies, and specific antibodies against ASFV to promote the development of prevention techniques are urgently needed. In the present study, pK205R of ASFV was successfully expressed in mammalian cells using a suspension culture system. An indirect enzyme-linked immunosorbent assay (ELISA) based on the purified pK205R was established and optimized. The monoclonal antibody (mAb) against pK205R recognized a conservative linear epitope (2VEPREQFFQDLLSAV16) and exhibited specific reactivity, which was conducive to the identification of the recombinant porcine reproductive and respiratory syndrome virus (PRRSV) expressing pK205R. The ELISA method efficiently detected clinical ASFV infection and revealed good application prospects in monitoring the antibody level in vivo for recombinant PRRSV live vector virus expressing the ASFV antigen protein. The determination of the conserved linear epitope of pK205R would contribute to further research on the structural biology and function of pK205R. The indirect ELISA method and mAb against ASFV pK205R revealed efficient detection and promising application prospects, making them ideal for epidemiological surveillance and vaccine research on ASF.African swine fever (ASF) is a contagious infectious disease with high lethality which continuously threatens the global pig industry causing huge economic losses. Currently, there are no commercially available vaccines or antiviral drugs that can effectively control ASF. The pathogen of ASF, ASF virus (ASFV) is a double-stranded DNA virus with a genome ranging from 170 to 193 kb and 151 to 167 open reading frames in various strains, which encodes 150-200 proteins. An effective method of monitoring ASFV antibodies, and specific antibodies against ASFV to promote the development of prevention techniques are urgently needed. In the present study, pK205R of ASFV was successfully expressed in mammalian cells using a suspension culture system. An indirect enzyme-linked immunosorbent assay (ELISA) based on the purified pK205R was established and optimized. The monoclonal antibody (mAb) against pK205R recognized a conservative linear epitope (2VEPREQFFQDLLSAV16) and exhibited specific reactivity, which was conducive to the identification of the recombinant porcine reproductive and respiratory syndrome virus (PRRSV) expressing pK205R. The ELISA method efficiently detected clinical ASFV infection and revealed good application prospects in monitoring the antibody level in vivo for recombinant PRRSV live vector virus expressing the ASFV antigen protein. The determination of the conserved linear epitope of pK205R would contribute to further research on the structural biology and function of pK205R. The indirect ELISA method and mAb against ASFV pK205R revealed efficient detection and promising application prospects, making them ideal for epidemiological surveillance and vaccine research on ASF. African swine fever (ASF) is a contagious infectious disease with high lethality which continuously threatens the global pig industry causing huge economic losses. Currently, there are no commercially available vaccines or antiviral drugs that can effectively control ASF. The pathogen of ASF, ASF virus (ASFV) is a double-stranded DNA virus with a genome ranging from 170 to 193 kb and 151 to 167 open reading frames in various strains, which encodes 150–200 proteins. An effective method of monitoring ASFV antibodies, and specific antibodies against ASFV to promote the development of prevention techniques are urgently needed. In the present study, pK205R of ASFV was successfully expressed in mammalian cells using a suspension culture system. An indirect enzyme-linked immunosorbent assay (ELISA) based on the purified pK205R was established and optimized. The monoclonal antibody (mAb) against pK205R recognized a conservative linear epitope (2VEPREQFFQDLLSAV16) and exhibited specific reactivity, which was conducive to the identification of the recombinant porcine reproductive and respiratory syndrome virus (PRRSV) expressing pK205R. The ELISA method efficiently detected clinical ASFV infection and revealed good application prospects in monitoring the antibody level in vivo for recombinant PRRSV live vector virus expressing the ASFV antigen protein. The determination of the conserved linear epitope of pK205R would contribute to further research on the structural biology and function of pK205R. The indirect ELISA method and mAb against ASFV pK205R revealed efficient detection and promising application prospects, making them ideal for epidemiological surveillance and vaccine research on ASF. |
| Author | Zhao, Ran Zhou, Yanjun Liu, Jiachen Li, Guoxin Guo, Ziqiang Tong, Wu Zheng, Haihong Qiao, Sina Liu, Changlong Gao, Fei Li, Liwei Dong, Shishan Jiang, Yifeng Tong, Guangzhi |
| AuthorAffiliation | 1 Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences , Shanghai , China 2 College of Veterinary Medicine, Hebei Agricultural University , Baoding , China 3 Xiamen Center for Animal Disease Control and Prevention , Xiamen , China 4 Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonose, Yangzhou University , Yangzhou , China |
| AuthorAffiliation_xml | – name: 1 Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences , Shanghai , China – name: 3 Xiamen Center for Animal Disease Control and Prevention , Xiamen , China – name: 4 Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonose, Yangzhou University , Yangzhou , China – name: 2 College of Veterinary Medicine, Hebei Agricultural University , Baoding , China |
| Author_xml | – sequence: 1 givenname: Liwei surname: Li fullname: Li, Liwei – sequence: 2 givenname: Sina surname: Qiao fullname: Qiao, Sina – sequence: 3 givenname: Jiachen surname: Liu fullname: Liu, Jiachen – sequence: 4 givenname: Yanjun surname: Zhou fullname: Zhou, Yanjun – sequence: 5 givenname: Wu surname: Tong fullname: Tong, Wu – sequence: 6 givenname: Shishan surname: Dong fullname: Dong, Shishan – sequence: 7 givenname: Changlong surname: Liu fullname: Liu, Changlong – sequence: 8 givenname: Yifeng surname: Jiang fullname: Jiang, Yifeng – sequence: 9 givenname: Ziqiang surname: Guo fullname: Guo, Ziqiang – sequence: 10 givenname: Haihong surname: Zheng fullname: Zheng, Haihong – sequence: 11 givenname: Ran surname: Zhao fullname: Zhao, Ran – sequence: 12 givenname: Guangzhi surname: Tong fullname: Tong, Guangzhi – sequence: 13 givenname: Guoxin surname: Li fullname: Li, Guoxin – sequence: 14 givenname: Fei surname: Gao fullname: Gao, Fei |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/36700212$$D View this record in MEDLINE/PubMed |
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| CitedBy_id | crossref_primary_10_1016_j_ijbiomac_2025_139701 crossref_primary_10_3389_fvets_2023_1289676 crossref_primary_10_3390_v15091939 crossref_primary_10_1016_j_ijbiomac_2023_127311 crossref_primary_10_1016_j_ijbiomac_2024_139169 crossref_primary_10_3390_v15102081 |
| Cites_doi | 10.1016/j.rvsc.2009.06.005 10.1016/j.biotechadv.2015.10.015 10.1080/22221751.2019.1590128 10.1099/vir.0.82857-0 10.1016/j.antiviral.2019.02.018 10.3390/v14092021 10.3390/vaccines8030531 10.3201/eid1412.080591 10.1016/j.virusres.2012.10.020 10.19958/j.cnki.cn31-2031/s.20220424.009 10.1016/j.vetmic.2020.108833 10.1515/pjvs-2016-0006 10.1128/JVI.01667-21 10.3390/v14020394 10.1371/journal.pone.0177007 10.3390/v13050943 10.1080/22221751.2021.1999779 10.1111/tbed.13132 10.1007/s11427-020-1657-9 10.1016/j.rvsc.2021.10.008 10.1080/22221751.2022.2108342 10.1111/tbed.12431 10.1007/s00253-022-11851-z 10.1128/jvi.00597-22 10.1128/CVI.00383-15 10.1099/0022-1317-83-6-1331 10.3389/fvets.2017.00081 10.1007/s00705-008-0246-z 10.3389/fvets.2021.781373 10.4238/2015.May.4.15 10.1016/j.vaccine.2020.10.057 10.1128/JVI.01199-21 10.3390/ani12141754 10.1007/s11427-021-1904-4 10.1080/22221751.2019.1565915 10.1016/j.vaccine.2018.04.079 10.3390/vaccines8040763 |
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| Copyright | Copyright © 2023 Li, Qiao, Liu, Zhou, Tong, Dong, Liu, Jiang, Guo, Zheng, Zhao, Tong, Li and Gao. Copyright © 2023 Li, Qiao, Liu, Zhou, Tong, Dong, Liu, Jiang, Guo, Zheng, Zhao, Tong, Li and Gao 2023 Li, Qiao, Liu, Zhou, Tong, Dong, Liu, Jiang, Guo, Zheng, Zhao, Tong, Li and Gao |
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| Keywords | recombinant PRRSV 293i cells indirect ELISA epitope ASFV pK205R |
| Language | English |
| License | Copyright © 2023 Li, Qiao, Liu, Zhou, Tong, Dong, Liu, Jiang, Guo, Zheng, Zhao, Tong, Li and Gao. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
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| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Edited by: Shaobin Shang, College of Veterinary Medicine, China Reviewed by: Yan-Dong Tang, Harbin Veterinary Research Institute (CAAS), China; Tao Lin, InnovHope, New Hope Group, United States These authors have contributed equally to this work This article was submitted to Viral Immunology, a section of the journal Frontiers in Immunology |
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| Snippet | African swine fever (ASF) is a contagious infectious disease with high lethality which continuously threatens the global pig industry causing huge economic... |
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| SubjectTerms | 293i cells African Swine Fever African Swine Fever Virus Animals Antibodies, Monoclonal - immunology ASFV pK205R Enzyme-Linked Immunosorbent Assay - methods epitope Immunology indirect ELISA recombinant PRRSV Swine Viral Proteins |
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| Title | A highly efficient indirect ELISA and monoclonal antibody established against African swine fever virus pK205R |
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