非洲猪瘟病毒pD345L蛋白单克隆抗体的制备及其表位鉴定

doi:10.3864/j.issn.0578-1752.2025.16.016

摘要/Abstract

摘要：

【目的】制备非洲猪瘟病毒（African swine fever virus，ASFV）pD345L蛋白（pD345L）的单克隆抗体（monoclonal antibodies, mAbs），并分析其识别的线性B细胞表位，为探究pD345L在ASFV感染与致病中的功能奠定基础。【方法】构建pET-28a-D345L重组原核表达质粒，将其转化大肠杆菌BL21（DE3）感受态细胞，表达并用亲和层析方法纯化重组的pD345L蛋白（rpD345L）。将纯化的rpD345L与等体积的弗氏完全佐剂乳化后免疫5周龄SPF级BALB/c小鼠，每隔两周免疫一次，共免疫3次。第二次和第三次免疫均用弗氏不完全佐剂乳化。第三次免疫一周后通过小鼠颌下静脉采血，分离血清，利用ELISA方法检测血清抗体效价，选取抗体效价高的小鼠加强免疫。免疫3 d后取小鼠脾细胞与小鼠骨髓瘤细胞（SP2/0）进行融合。将重组GST-pD345L作为包被抗原，利用ELISA方法筛选阳性杂交瘤细胞，经过3次筛选得到能够稳定分泌pD345L mAbs的杂交瘤细胞株，注入小鼠腹腔制备腹水。使用单抗亚类鉴定试剂盒鉴定mAbs的重、轻链类型。利用外源过表达的pD345L和ASFV HLJ/18株感染的猪肺泡巨噬细胞（PAMs）为抗原，用制备的mAbs作为一抗，分别进行Western blot和间接免疫荧光（IFA）鉴定。随后将截短的pD345L与GST融合表达，用筛选的mAbs鉴定抗原表位，分析鉴定的抗原表位在不同ASFV毒株中的保守性。【结果】IPTG诱导后，pD345L以包涵体形式表达，分子量为40.5 kDa。将其免疫BALB/c小鼠后经间接ELISA方法筛选得到2株稳定分泌pD345L mAbs的杂交瘤细胞株，命名为10H3和5G1。亚类鉴定结果显示，2株mAbs重链类型均为IgG1型，轻链类型为κ链，抗体效价为1:1 638 400。Western blot和IFA结果显示，制备的mAbs均能识别天然表达的pD345L。经Western blot确定抗原表位，结果显示10H3和5G1 mAbs的最小线性B细胞表位分别为¹METFVRLFKD¹⁰和³²¹YEKICCSEES³³⁰，这两个抗原表位在ASFV不同毒株中具有保守性。【结论】本研究原核表达并纯化了rpD345L，制备了2株pD345L mAbs，并鉴定了其识别的抗原表位，且这2株mAbs均能有效识别ASFV感染过程中表达的pD345L，为进一步研究ASFV pD345L功能奠定基础。

关键词: 非洲猪瘟病毒, pD345L蛋白, 单克隆抗体, 抗原表位

Abstract:

【Objective】The aim of this study was to prepare monoclonal antibodies (mAbs) against the pD345L protein (pD345L) of African swine fever virus (ASFV), and to analyze the linear B-cell epitopes recognized by these antibodies, so as to lay a foundation for exploring the function of pD345L in the infection and pathogenesis of ASFV. 【Method】The prokaryotic expression plasmid pET-28a-D345L was constructed, transformed into Escherichia coli BL21(DE3) receptor cells, expressed and purified recombinant pD345L protein (rpD345L) by affinity chromatography. The purified recombinant pD345L protein was emulsified with an equal volume of Freund's complete adjuvant and immunized 5-week-old SPF BALB/c mice once every two weeks for a total of 3 times. Both the second and third immunization were emulsified with Freund's incomplete adjuvant. A week after the third immunization, the blood sample was collected through the submaxillary vein of mice, serum was separated, serum antibody titer was detected by indirect enzyme-linked immunosorbent assay (ELISA), and mice with high antibody titer were selected to enhance immunity. After 3 days of immunization, the mouse spleen cells were fused with mouse myeloma cells (SP2/0). Recombinant GST-pD345L was used as the coated antigen, and positive hybridoma cells were screened by ELISA. After three-time screening, hybridoma cell lines that could secrete pD345L protein mAbs were obtained and injected into the abdominal cavity of mice to prepare ascites. The heavy and light chain types of mAbs were identified using the monoclonal antibody subclass identification kit. The overexpressed pD345L protein and ASFV HLJ/18 infected porcine alveolar macrophages (PAMs) were identified by Western blot and indirect immunofluorescence (IFA), respectively, using the prepared mAbs as the primary antibody. Then the truncated pD345L protein was fused with GST, and the epitopes were identified with screened mAbs, and the conserved types of identified epitopes in different ASFV strains were analyzed. 【Result】After induction by IPTG, pD345L was expressed as an inclusion body with a molecular weight of 40.5 kDa. After immunizing BALB/c mice, two hybridoma cell lines with stable secretion of pD345L mAbs were selected by indirect ELISA method, and named 10H3 and 5G1. The results of subclass identification showed that the heavy chain type of the two strains of mAbs was IgG1 type, the light chain type was κ chain, and the antibody titer was 1:1 638 400. Western blot and IFA results showed that the prepared mAbs could recognize naturally expressed pD345L. The results showed that the minimum linear B-cell epitopes of 10H3 and 5G1 mAbs were ¹METFVRLFKD¹⁰ and ³²¹YEKICCSEES³³⁰, respectively. These two epitopes were conserved in different ASFV strains. 【Conclusion】In this study, the recombinant pD345L protein was expressed and purified in prokaryotes, two strains of pD345L protein mAbs were prepared, and their recognized epitopes were identified. Both strains of mAbs could effectively recognize pD345L protein expressed during ASFV infection, which laid a foundation for further research on the function of ASFV pD345L protein.

Key words: African swine fever virus, pD345L protein, monoclonal antibodies, epitope

贺筱萍, 张元峰, 刘学敏, 黄丽, 翁长江. 非洲猪瘟病毒pD345L蛋白单克隆抗体的制备及其表位鉴定[J]. 中国农业科学, 2025, 58(16): 3345-3356.

HE XiaoPing, ZHANG YuanFeng, LIU XueMin, HUANG Li, WENG ChangJiang. Preparation of Monoclonal Antibody Against African Swine Fever Virus pD345L Protein and Identification of Its Epitope[J]. Scientia Agricultura Sinica, 2025, 58(16): 3345-3356.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2025.16.016

https://www.chinaagrisci.com/CN/Y2025/V58/I16/3345

图/表 7

表1

ASFV D345L基因及其截短片段的PCR扩增引物"

片段Fragment	引物 Primers (5'-3')
D345L D345L-D1	F: 5'-ATGGGTCGCGGATCC GAATTCATGGAAACCTTTGTACGC-3' R: 5'-CACCACCACCACCAC CTCGAGCTAAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3'
D345L-D2	R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGTA-3' F: 5'-TTCCAGGGGCCCCTG GGATCCCTAACGCTGGGATACACT-3'
D345L-D3 D345L-P4 D345L-P5 D345L-P6 D345L-P7 D345L-P8 D345L-P9 D345L-P10 D345L-P11 D345L-P12 D345L-P13 D345L-P14 D345L-P15 D345L-P16	R: 5'-GTCACGATGCGGCCG CTCGAGGCCAAAATCTTGAGGAGC-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCCT-3' R: 5'-GTCACGATGCGGCCG CTCGAGCTAAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3' R: 5'-GTCACGATGCGGCCG CTCGAGGTGACAAAACCCATCGG-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAAACCCATCGGGGCTAT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3' R: 5'-GTCACGATGCGGCCG CTCGAGGTATCCCAAATAGGTCC-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3' R: 5'-GTCACGATGCGGCCG CTCGAGATTGTCACCTATGACG-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3' R: 5'-GTCACGATGCGGCCG CTCGAGCAACTCAAGCAGATCTTT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAAAAAGGGTTCCCCAGC-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3' R: 5'-GTCACGATGCGGCCG CTCGAGGTTCAAATTCTTCTTAA-3' F: 5'- TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3' R: 5'-GTCACGATGCGGCCG CTCGAGTTCCGCCAAGGTTATAT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGGGTTTAAACCTAAA-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGAGCCACCCACCTGAGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3' R: 5'-GTCACGATGCGGCCG CTCGAG ATGCCAGGCATCGGAGC-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATGGAAACCTTTGTACGC-3' R: 5'-GTCACGATGCGGCCG CTCGAGGTCTTTAAACAGGCGTACA-3' F: 5'-TTCCAGGGGCCCCTG GGATCCTCTCCTCAGCAGCGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGT-3'
D345L-P17 D345L-P18 D345L-P19 D345L-P20 D345L-P21 D345L-P22 D345L-P23 D345L-P24 D345L-P25 D345L-P26 D345L-P27 D345L-P28 D345L-P29 D345L-P30 D345L-P31 D345L-P32 D345L-P33 D345L-P34 D345L-P35 D345L-P36 D345L-P37 D345L-P38	F: 5'-TTCCAGGGGCCCCTG GGATCCGCTATTCGTCGCACTCAG-3' R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGACCTTGCCAGCGTTTTA-3' R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCTACAAAAGCTATTATATA-3' R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCAAAGCAAATCTTTTTAA-3' R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCCGCGCTGCTTGTAGCTG-3' R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCGTGTTAGTAAAGA-3' R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCTTCTGCCAAACCACCGTC-3' R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATTCATATTGATGGGACC-3' R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCCCCGGACATAGTAATAGC-3' R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCTGTCACCTAACGCTGGGA-3' R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCCTAACGCTGGGATACACT-3' R: 5'-GTCACGATGCGGCCG CTCGAGCCAGGACTGTTGAGTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAAAATTGTCGATGTCCTC-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAAAATTGTCGATGTCCTC-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAGCATCTCCATGGGGGATG-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTATGTAGGTCACGTA-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAGGATGGGATATTTGTCTTT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAGATGTTTCATAACTGGAATT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAAAGACCTTCCATCCAA-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAGATTTGGCATTTCGGGACA-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGGCATAGAATAGTGAA-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAGTTTTTGATACAGCGTTTCA-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAAATGTTTCCGCGTCGAGCG-3'
D345L-P39 D345L-P40 D345L-P41 D345L-P42 D345L-P43 D345L-P44 D345L-P45 D345L-P46 D345L-P47 D345L-P48 D345L-P49 D345L-P50 D345L-P51	F: 5'-TTCCAGGGGCCCCTG GGATCCGAGCACACCGAGGCTCC-3' R: 5'-GTCACGATGCGGCCG CTCGAG TTGAGGAGCCTCGGTGTGC-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGATTTTGGCACGCTCGAC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCCGCCAACTACTTGAAA-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGATCAGTACACTATTCA-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCTATGAAACCGCGTGTC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGTGGTTGGCTACTTTG-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATCTTTCAAATAATTCCA-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCCCCCAGTTTTTAAAAGA-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATACAACAGTTCTTACGT-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCATTAAAGCCTCGCCATC-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCTATGAAAAAATTTGCTG-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCGCTCTATCCACAGAGG-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTAAGCATATCTGT-3' F: 5'-TTCCAGGGGCCCCTG GGATCCACAGATATGCTTACT-3' R: 5'-GTCACGATGCGGCCG CTCGAGAGTAAGCATATCTGT-3'

表1

图1

图2

图3

图4

图5

图6

参考文献 30

[1]	SÁNCHEZ-CORDÓN P J, MONTOYA M, REIS A L, DIXON L K. African swine fever: A re-emerging viral disease threatening the global pig industry. The Veterinary Journal, 2018, 233: 41-48.
[2]	LI Z Y, CHEN W X, QIU Z L, LI Y W, FAN J D, WU K K, LI X W, ZHAO M Q, DING H X, FAN S Q, CHEN J D. African swine fever virus: A review. Life, 2022, 12(8): 1255.
[3]	DIXON L K, SUN H, ROBERTS H. African swine fever. Antiviral Research, 2019, 165: 34-41. doi: S0166-3542(19)30096-8 pmid: 30836106
[4]	SÁNCHEZ-VIZCAÍNO J M, MUR L, MARTÍNEZ-LÓPEZ B. African swine fever (ASF): Five years around Europe. Veterinary Microbiology, 2013, 165(1/2): 45-50.
[5]	冯春莹, 张朝霞, 刘云飞, 黄丽, 翁长江. 非洲猪瘟病毒p54蛋白单克隆抗体的制备及其抗原表位鉴定. 中国农业科学, 2024, 57(19): 3936-3944. doi: 10.3864/j.issn.0578-1752.2024.19.015.
	FENG C Y, ZHANG Z X, LIU Y F, HUANG L, WENG C J. Preparation of monoclonal antibody againstafrican swine fever virus P54 protein and identification of its epitope. Scientia Agricultura Sinica, 2024, 57(19): 3936-3944. doi: 10.3864/j.issn.0578-1752.2024.19.015. (in Chinese)
[6]	罗玉子, 孙元, 王涛, 仇华吉. 非洲猪瘟——我国养猪业的重大威胁. 中国农业科学, 2018, 51(21): 4177-4187. doi: 10.3864/j.issn.0578-1752.2018.21.016.
	LUO Y Z, SUN Y, WANG T, QIU H J. African swine fever: A major threat to the Chinese swine industry. Scientia Agricultura Sinica, 2018, 51(21): 4177-4187. doi: 10.3864/j.issn.0578-1752.2018.21.016. (in Chinese)
[7]	DIXON L K, CHAPMAN D A G, NETHERTON C L, UPTON C. African swine fever virus replication and genomics. Virus Research, 2013, 173(1): 3-14. doi: 10.1016/j.virusres.2012.10.020 pmid: 23142553
[8]	REVILLA Y, PÉREZ-NÚÑEZ D, RICHT J A. African swine fever virus biology and vaccine approaches. Advances in Virus Research. Amsterdam: Elsevier, 2018: 41-74.
[9]	WANG Y, KANG W F, YANG W P, ZHANG J, LI D, ZHENG H X. Structure of African swine fever virus and associated molecular mechanisms underlying infection and immunosuppression: A review. Frontiers in Immunology, 2021, 12: 715582.
[10]	WANG G G, XIE M J, WU W, CHEN Z Z. Structures and functional diversities of ASFV proteins. Viruses, 2021, 13(11): 2124.
[11]	CHEN H, WANG Z Z, GAO X Y, LV J X, HU Y X, JUNG Y S, ZHU S Y, WU X D, QIAN Y J, DAI J J. ASFV pD345L protein negatively regulates NF-κB signalling by inhibiting IKK kinase activity. Veterinary Research, 2022, 53(1): 32. doi: 10.1186/s13567-022-01050-z pmid: 35461299
[12]	WANG Z Y, AI Q Y, HUANG S L, OU Y T, GAO Y Z, TONG T Z, FAN H Y. Immune escape mechanism and vaccine research progress of African swine fever virus. Vaccines, 2022, 10(3): 344.
[13]	LI L, FU J Y, LI J X, GUO S B, CHEN Q C, ZHANG Y B, LIU Z K, TAN C, CHEN H C, WANG X R. African swine fever virus pI215L inhibits type I interferon signaling by targeting interferon regulatory factor 9 for autophagic degradation. Journal of Virology, 2022, 96(17): e0094422.
[14]	LIU H S, ZHU Z X, FENG T, MA Z, XUE Q, WU P X, LI P, LI S S, YANG F, CAO W J, XUE Z N, CHEN H J, LIU X T, ZHENG H X. African swine fever virus E120R protein inhibits interferon beta production by interacting with IRF3 to block its activation. Journal of Virology, 2021, 95(18): e0082421.
[15]	CHEN S, ZHANG X H, NIE Y, LI H X, CHEN W G, LIN W C, CHEN F, XIE Q M. African swine fever virus protein E199L promotes cell autophagy through the interaction of PYCR2. Virologica Sinica, 2021, 36(2): 196-206. doi: 10.1007/s12250-021-00375-x pmid: 33830435
[16]	WANG T, LUO R, ZHANG J, LU Z H, LI L F, ZHENG Y H, PAN L, LAN J, ZHAI H J, HUANG S J, SUN Y, QIU H J. The MGF300-2R protein of African swine fever virus is associated with viral pathogenicity by promoting the autophagic degradation of IKKα and IKKβ through the recruitment of TOLLIP. PLoS Pathogens, 2023, 19(8): e1011580.
[17]	GAO Q, YANG Y L, LUO Y Z, CHEN X N, GONG T, WU D D, FENG Y Z, ZHENG X Y, WANG H, ZHANG G H, LU G, GONG L. African swine fever virus envelope glycoprotein CD2v interacts with host CSF2RA to regulate the JAK2-STAT3 pathway and inhibit apoptosis to facilitate virus replication. Journal of Virology, 2023, 97(4): e0188922.
[18]	TRAN X H, LE T T P, NGUYEN Q H, DO T T, NGUYEN V D, GAY C G, BORCA M V, GLADUE D P. African swine fever virus vaccine candidate ASFV-G-ΔI177L efficiently protects European and native pig breeds against circulating Vietnamese field strain. Transboundary and Emerging Diseases, 2022, 69(4): e497-e504.
[19]	王涛, 罗瑞, 孙元, 仇华吉. 非洲猪瘟疫苗的研发策略与应用前景:可行性和可能性. 中国农业科学, 2023, 56(11): 2212-2222. doi: 10.3864/j.issn.0578-1752.2023.11.014.
	WANG T, LUO R, SUN Y, QIU H J. Development strategies and application prospects of African swine fever vaccines: Feasibility and Probability. Scientia Agricultura Sinica, 2023, 56(11): 2212-2222. doi: 10.3864/j.issn.0578-1752.2023.11.014. (in Chinese)
[20]	BORCA M V, RAMIREZ-MEDINA E, SILVA E, RAI A, ESPINOZA N, VELAZQUEZ-SALINAS L, GLADUE D P. ASF vaccine candidate ASFV-G-∆I177L does not exhibit residual virulence in long- term clinical studies. Pathogens, 2023, 12(6): 805.
[21]	CHATHURANGA K, LEE J S. African swine fever virus (ASFV): immunity and vaccine development. Vaccines, 2023, 11(2): 199.
[22]	SONG J X, WANG M X, ZHOU L, TIAN P P, SUN Z Y, SUN J R, WANG X N, ZHUANG G Q, JIANG D W, WU Y N, ZHANG G P. A candidate nanoparticle vaccine comprised of multiple epitopes of the African swine fever virus elicits a robust immune response. Journal of Nanobiotechnology, 2023, 21(1): 424. doi: 10.1186/s12951-023-02210-9 pmid: 37964304
[23]	HUANG L, CHEN W Y, LIU H Y, XUE M D, DONG S Q, LIU X H, FENG C Y, CAO S N, YE G Q, ZHOU Q Q, et al. African swine fever virus HLJ/ 18 CD2v suppresses type I IFN production and IFN- stimulated genes expression through negatively regulating cGMP- AMP synthase-STING and IFN signaling pathways. The Journal of Immunology, 2023, 210(9): 1338-1350.
[24]	LIU X H, CHEN H F, YE G Q, LIU H Y, FENG C Y, CHEN W Y, HU L, ZHOU Q Q, ZHANG Z X, LI J N, et al. African swine fever virus pB318L, a trans-geranylgeranyl-diphosphate synthase, negatively regulates cGAS-STING and IFNAR-JAK-STAT signaling pathways. PLoS Pathogens, 2024, 20(4): e1012136.
[25]	YE G Q, LIU H Y, LIU X H, CHEN W Y, LI J N, ZHAO D M, WANG G, FENG C Y, ZHANG Z X, ZHOU Q Q, et al. African swine fever virus H240R protein inhibits the production of type I interferon through disrupting the oligomerization of STING. Journal of Virology, 2023, 97(9): e0057723.
[26]	ANTONIA R J, HAGAN R S, BALDWIN A S. Expanding the view of IKK: New substrates and new biology. Trends in Cell Biology, 2021, 31(3): 166-178. doi: 10.1016/j.tcb.2020.12.003 pmid: 33422358
[27]	TAMIMI A, TAMIMI A, SORKHEH F, ASL S M, GHAFARI A, KARIMI A G, ERABI G, POURMONTASERI H, DERAVI N. Monoclonal antibodies for the treatment of squamous cell carcinoma: A literature review. Cancer Reports, 2023, 6(5): e1802.
[28]	JIN J, SIMMONS G. Antiviral functions of monoclonal antibodies against chikungunya virus. Viruses, 2019, 11(4): 305.
[29]	XIANG R, WANG Y, WANG L L, DENG X Q, HUO S S, JIANG S B, YU F. Neutralizing monoclonal antibodies against highly pathogenic coronaviruses. Current Opinion in Virology, 2022, 53: 101199.
[30]	TSAO L C, FORCE J, HARTMAN Z C. Mechanisms of therapeutic antitumor monoclonal antibodies. Cancer Research, 2021, 81(18): 4641-4651.