Identification and characterization of nanobodies specifically against African swine fever virus major capsid protein p72
African swine fever virus (ASFV) is a large and very complex DNA virus. The major capsid protein p72 is the most predominant structural protein and constitutes the outmost icosahedral capsid of the virion. In the present study, the nanobodies against ASFV p72 protein were screened from a camelid imm...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
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Frontiers Media S.A.
2022-10-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2022.1017792/full |
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| author | Jifei Yang Mengyao Jing Qingli Niu Jinming Wang Yaru Zhao Yaru Zhao Meng Liu Guiquan Guan Jianxun Luo Hong Yin Hong Yin Zhijie Liu Zhijie Liu |
| author_facet | Jifei Yang Mengyao Jing Qingli Niu Jinming Wang Yaru Zhao Yaru Zhao Meng Liu Guiquan Guan Jianxun Luo Hong Yin Hong Yin Zhijie Liu Zhijie Liu |
| author_sort | Jifei Yang |
| collection | DOAJ |
| description | African swine fever virus (ASFV) is a large and very complex DNA virus. The major capsid protein p72 is the most predominant structural protein and constitutes the outmost icosahedral capsid of the virion. In the present study, the nanobodies against ASFV p72 protein were screened from a camelid immune VHH library by phage display technique. Nine distinct nanobodies were identified according to the amino acid sequences of the complementary determining regions (CDRs), and contain typical amino acid substitutions in the framework region 2 (FR2). Six nanobodies were successfully expressed in E. coli, and their specificity and affinity to p72 protein were further evaluated. The results showed that nanobodies Nb25 had the best affinity to both recombinant and native p72 protein of ASFV. The Nb25 possesses an extremely long CDR3 with 23 amino acids compared with other nanobodies, which may allow this nanobody to access the hidden epitopes of target antigen. Furthermore, the Nb25 can specifically recognize the virus particles captured by polyclonal antibody against ASFV in a sandwich immunoassay, and its application as a biosensor to target virus in PAM cells was verified by an immunofluorescence assay. Nanobodies have been proven to possess many favorable properties with small size, high affinity and specificity, easier to produce, low costs and deep tissue penetration that make them suitable for various biotechnological applications. These findings suggest that nanobody Nb25 identified herein could be a valuable alternative tool and has potential applications in diagnostic and basic research on ASFV. |
| first_indexed | 2024-04-11T17:21:01Z |
| format | Article |
| id | doaj.art-1add670808cc482f9e8bafcfa94971b9 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-11T17:21:01Z |
| publishDate | 2022-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-1add670808cc482f9e8bafcfa94971b92022-12-22T04:12:30ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2022-10-011310.3389/fmicb.2022.10177921017792Identification and characterization of nanobodies specifically against African swine fever virus major capsid protein p72Jifei Yang0Mengyao Jing1Qingli Niu2Jinming Wang3Yaru Zhao4Yaru Zhao5Meng Liu6Guiquan Guan7Jianxun Luo8Hong Yin9Hong Yin10Zhijie Liu11Zhijie Liu12African Swine Fever Regional Laboratory of China (Lanzhou), State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, ChinaAfrican Swine Fever Regional Laboratory of China (Lanzhou), State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, ChinaAfrican Swine Fever Regional Laboratory of China (Lanzhou), State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, ChinaAfrican Swine Fever Regional Laboratory of China (Lanzhou), State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, ChinaAfrican Swine Fever Regional Laboratory of China (Lanzhou), State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, ChinaChina Agricultural VET. BIO. Science and Technology Co, Ltd, Lanzhou, ChinaAnimal Husbandry and Veterinary Bureau of Dingxi City, Dingxi, Gansu, ChinaAfrican Swine Fever Regional Laboratory of China (Lanzhou), State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, ChinaAfrican Swine Fever Regional Laboratory of China (Lanzhou), State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, ChinaAfrican Swine Fever Regional Laboratory of China (Lanzhou), State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, ChinaJiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonosis, Yangzhou University, Yangzhou, ChinaAfrican Swine Fever Regional Laboratory of China (Lanzhou), State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, ChinaInstitute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, ChinaAfrican swine fever virus (ASFV) is a large and very complex DNA virus. The major capsid protein p72 is the most predominant structural protein and constitutes the outmost icosahedral capsid of the virion. In the present study, the nanobodies against ASFV p72 protein were screened from a camelid immune VHH library by phage display technique. Nine distinct nanobodies were identified according to the amino acid sequences of the complementary determining regions (CDRs), and contain typical amino acid substitutions in the framework region 2 (FR2). Six nanobodies were successfully expressed in E. coli, and their specificity and affinity to p72 protein were further evaluated. The results showed that nanobodies Nb25 had the best affinity to both recombinant and native p72 protein of ASFV. The Nb25 possesses an extremely long CDR3 with 23 amino acids compared with other nanobodies, which may allow this nanobody to access the hidden epitopes of target antigen. Furthermore, the Nb25 can specifically recognize the virus particles captured by polyclonal antibody against ASFV in a sandwich immunoassay, and its application as a biosensor to target virus in PAM cells was verified by an immunofluorescence assay. Nanobodies have been proven to possess many favorable properties with small size, high affinity and specificity, easier to produce, low costs and deep tissue penetration that make them suitable for various biotechnological applications. These findings suggest that nanobody Nb25 identified herein could be a valuable alternative tool and has potential applications in diagnostic and basic research on ASFV.https://www.frontiersin.org/articles/10.3389/fmicb.2022.1017792/fullAfrican swine fevermajor capsid protein p72phage displaynanobodybiosensor |
| spellingShingle | Jifei Yang Mengyao Jing Qingli Niu Jinming Wang Yaru Zhao Yaru Zhao Meng Liu Guiquan Guan Jianxun Luo Hong Yin Hong Yin Zhijie Liu Zhijie Liu Identification and characterization of nanobodies specifically against African swine fever virus major capsid protein p72 Frontiers in Microbiology African swine fever major capsid protein p72 phage display nanobody biosensor |
| title | Identification and characterization of nanobodies specifically against African swine fever virus major capsid protein p72 |
| title_full | Identification and characterization of nanobodies specifically against African swine fever virus major capsid protein p72 |
| title_fullStr | Identification and characterization of nanobodies specifically against African swine fever virus major capsid protein p72 |
| title_full_unstemmed | Identification and characterization of nanobodies specifically against African swine fever virus major capsid protein p72 |
| title_short | Identification and characterization of nanobodies specifically against African swine fever virus major capsid protein p72 |
| title_sort | identification and characterization of nanobodies specifically against african swine fever virus major capsid protein p72 |
| topic | African swine fever major capsid protein p72 phage display nanobody biosensor |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2022.1017792/full |
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