Preparation and epitope mapping of monoclonal antibodies against African swine fever virus p22 protein
Copyright © 2023. Published by Elsevier B.V..
African swine fever (ASF), caused by the African swine fever virus (ASFV), is now widespread in many countries and severely affects the commercial rearing of swine. Rapid and early diagnosis is crucial for the prevention of ASF. ASFV mature virions comprise the inner envelope protein, p22, making it an excellent candidate for the serological diagnosis and surveillance of ASF. In this study, the prokaryotic-expressed p22 recombinant protein was prepared and purified for immunization in mice. Four monoclonal antibodies (mAbs) were identified using hybridoma cell fusion, clone purification, and immunological assays. The epitopes of mAbs 14G1 and 22D8 were further defined by alanine-scanning mutagenesis. Our results showed that amino acids C39, K40, V41, D42, C45, G48, E49, and C51 directly bound to 14G1, while the key amino acid epitope for 22D8 included K161, Y162, G163, D165, H166, I167, and I168. Homologous and structural analysis revealed that these sites were highly conserved across Asian and European ASFV strains, and the amino acids identified were located on the surface of p22. Thus, our study contributes to a better understanding of the antigenicity of the ASFV p22 protein, and the results could facilitate the prevention and control of ASF.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2024 |
|---|---|
| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:255 |
|---|---|
| Enthalten in: |
International journal of biological macromolecules - 255(2024) vom: 07. Jan., Seite 128111 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Shi, Lan-Fang [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
African swine fever virus |
|---|
| Anmerkungen: |
Date Completed 10.01.2024 Date Revised 10.01.2024 published: Print-Electronic Citation Status MEDLINE |
|---|
| doi: |
10.1016/j.ijbiomac.2023.128111 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM364712767 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM364712767 | ||
| 003 | DE-627 | ||
| 005 | 20240114233421.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231226s2024 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1016/j.ijbiomac.2023.128111 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1255.xml |
| 035 | |a (DE-627)NLM364712767 | ||
| 035 | |a (NLM)37979744 | ||
| 035 | |a (PII)S0141-8130(23)05010-9 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Shi, Lan-Fang |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Preparation and epitope mapping of monoclonal antibodies against African swine fever virus p22 protein |
| 264 | 1 | |c 2024 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 10.01.2024 | ||
| 500 | |a Date Revised 10.01.2024 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2023. Published by Elsevier B.V. | ||
| 520 | |a African swine fever (ASF), caused by the African swine fever virus (ASFV), is now widespread in many countries and severely affects the commercial rearing of swine. Rapid and early diagnosis is crucial for the prevention of ASF. ASFV mature virions comprise the inner envelope protein, p22, making it an excellent candidate for the serological diagnosis and surveillance of ASF. In this study, the prokaryotic-expressed p22 recombinant protein was prepared and purified for immunization in mice. Four monoclonal antibodies (mAbs) were identified using hybridoma cell fusion, clone purification, and immunological assays. The epitopes of mAbs 14G1 and 22D8 were further defined by alanine-scanning mutagenesis. Our results showed that amino acids C39, K40, V41, D42, C45, G48, E49, and C51 directly bound to 14G1, while the key amino acid epitope for 22D8 included K161, Y162, G163, D165, H166, I167, and I168. Homologous and structural analysis revealed that these sites were highly conserved across Asian and European ASFV strains, and the amino acids identified were located on the surface of p22. Thus, our study contributes to a better understanding of the antigenicity of the ASFV p22 protein, and the results could facilitate the prevention and control of ASF | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a African swine fever virus | |
| 650 | 4 | |a Epitope mapping | |
| 650 | 4 | |a Monoclonal antibodies | |
| 650 | 4 | |a p22 | |
| 650 | 7 | |a Antibodies, Monoclonal |2 NLM | |
| 650 | 7 | |a Antibodies, Viral |2 NLM | |
| 650 | 7 | |a Epitopes |2 NLM | |
| 650 | 7 | |a Amino Acids |2 NLM | |
| 700 | 1 | |a Ren, Haojie |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Biao |e verfasserin |4 aut | |
| 700 | 1 | |a Shi, Sai-Yan |e verfasserin |4 aut | |
| 700 | 1 | |a Shao, Han-Cheng |e verfasserin |4 aut | |
| 700 | 1 | |a Xing, Hainan |e verfasserin |4 aut | |
| 700 | 1 | |a Zhao, Yan-Yan |e verfasserin |4 aut | |
| 700 | 1 | |a Lin, Zhi-Zhao |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Yuhang |e verfasserin |4 aut | |
| 700 | 1 | |a Han, Shichong |e verfasserin |4 aut | |
| 700 | 1 | |a He, Wen-Rui |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Gaiping |e verfasserin |4 aut | |
| 700 | 1 | |a Wan, Bo |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t International journal of biological macromolecules |d 1992 |g 255(2024) vom: 07. Jan., Seite 128111 |w (DE-627)NLM012627356 |x 1879-0003 |7 nnns |
| 773 | 1 | 8 | |g volume:255 |g year:2024 |g day:07 |g month:01 |g pages:128111 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1016/j.ijbiomac.2023.128111 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 255 |j 2024 |b 07 |c 01 |h 128111 | ||