Characterization of monoclonal antibodies against porcine epidemic diarrhea virus S1/S2 junction protein
Abstract Pig producers have faced considerable economic losses due to porcine epidemic diarrhea virus (PEDV) infection, emphasizing the need for PEDV antibody development. The S1/S2 junction (S1S2J) cleavage site of the S protein of PEDV is one of the major determinants of coronavirus infection succ...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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SpringerOpen
2023-07-01
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| Series: | AMB Express |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13568-023-01573-4 |
| _version_ | 1797778718284840960 |
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| author | Nan Huang Qiaoli Lang Liping Li Liangpeng Ge Xi Yang |
| author_facet | Nan Huang Qiaoli Lang Liping Li Liangpeng Ge Xi Yang |
| author_sort | Nan Huang |
| collection | DOAJ |
| description | Abstract Pig producers have faced considerable economic losses due to porcine epidemic diarrhea virus (PEDV) infection, emphasizing the need for PEDV antibody development. The S1/S2 junction (S1S2J) cleavage site of the S protein of PEDV is one of the major determinants of coronavirus infection success. In this study, we specifically selected the S1S2J protein of PEDV-AJ1102 (a representative strain of the G2 type) as a target protein to immunize mice and generated monoclonal antibodies (mAbs) using hybridoma technology. Three mAbs with high-binding activities to the S1S2J protein and were obtained and further analyzed. To reveal the characterization of these mAbs, variable region genes of antibodies were studied by using DNA sequencing, thereby revealing differences in their CDR3 amino acid sequences. We then developed a new method to identify the isotypes of these three mAbs. Results showed that these three antibodies were of the IgM type. As for the functions of these three mAbs, indirect immunofluorescence assay confirmed their good binding ability to Vero E6 cells infected with the PEDV-SP-C strain (G1 type). Epitope analysis showed linear epitopes for all three mAbs. These antibodies were also used to detect infected cells via flow cytometry analysis. In summary, we prepared and examined three mAbs against PEDV-S1S2J. These mAbs can be employed as detection antibodies for diagnostic reagents and further developed for other applications. We also designed a novel technique for easy and cost-saving identification of isotypes of mouse mAbs. Our results lay a good foundation for the development of research on PEDV. |
| first_indexed | 2024-03-12T23:20:31Z |
| format | Article |
| id | doaj.art-a80b4accdae04779bbfd03be92d43488 |
| institution | Directory Open Access Journal |
| issn | 2191-0855 |
| language | English |
| last_indexed | 2024-03-12T23:20:31Z |
| publishDate | 2023-07-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | AMB Express |
| spelling | doaj.art-a80b4accdae04779bbfd03be92d434882023-07-16T11:29:01ZengSpringerOpenAMB Express2191-08552023-07-011311910.1186/s13568-023-01573-4Characterization of monoclonal antibodies against porcine epidemic diarrhea virus S1/S2 junction proteinNan Huang0Qiaoli Lang1Liping Li2Liangpeng Ge3Xi Yang4Chongqing Academy of Animal SciencesChongqing Academy of Animal SciencesChongqing Academy of Animal SciencesChongqing Academy of Animal SciencesChongqing Academy of Animal SciencesAbstract Pig producers have faced considerable economic losses due to porcine epidemic diarrhea virus (PEDV) infection, emphasizing the need for PEDV antibody development. The S1/S2 junction (S1S2J) cleavage site of the S protein of PEDV is one of the major determinants of coronavirus infection success. In this study, we specifically selected the S1S2J protein of PEDV-AJ1102 (a representative strain of the G2 type) as a target protein to immunize mice and generated monoclonal antibodies (mAbs) using hybridoma technology. Three mAbs with high-binding activities to the S1S2J protein and were obtained and further analyzed. To reveal the characterization of these mAbs, variable region genes of antibodies were studied by using DNA sequencing, thereby revealing differences in their CDR3 amino acid sequences. We then developed a new method to identify the isotypes of these three mAbs. Results showed that these three antibodies were of the IgM type. As for the functions of these three mAbs, indirect immunofluorescence assay confirmed their good binding ability to Vero E6 cells infected with the PEDV-SP-C strain (G1 type). Epitope analysis showed linear epitopes for all three mAbs. These antibodies were also used to detect infected cells via flow cytometry analysis. In summary, we prepared and examined three mAbs against PEDV-S1S2J. These mAbs can be employed as detection antibodies for diagnostic reagents and further developed for other applications. We also designed a novel technique for easy and cost-saving identification of isotypes of mouse mAbs. Our results lay a good foundation for the development of research on PEDV.https://doi.org/10.1186/s13568-023-01573-4Porcine epidemic diarrhea virusMonoclonal antibodiesAntibody subclassesIsotype identification method |
| spellingShingle | Nan Huang Qiaoli Lang Liping Li Liangpeng Ge Xi Yang Characterization of monoclonal antibodies against porcine epidemic diarrhea virus S1/S2 junction protein AMB Express Porcine epidemic diarrhea virus Monoclonal antibodies Antibody subclasses Isotype identification method |
| title | Characterization of monoclonal antibodies against porcine epidemic diarrhea virus S1/S2 junction protein |
| title_full | Characterization of monoclonal antibodies against porcine epidemic diarrhea virus S1/S2 junction protein |
| title_fullStr | Characterization of monoclonal antibodies against porcine epidemic diarrhea virus S1/S2 junction protein |
| title_full_unstemmed | Characterization of monoclonal antibodies against porcine epidemic diarrhea virus S1/S2 junction protein |
| title_short | Characterization of monoclonal antibodies against porcine epidemic diarrhea virus S1/S2 junction protein |
| title_sort | characterization of monoclonal antibodies against porcine epidemic diarrhea virus s1 s2 junction protein |
| topic | Porcine epidemic diarrhea virus Monoclonal antibodies Antibody subclasses Isotype identification method |
| url | https://doi.org/10.1186/s13568-023-01573-4 |
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