Preparation of the monoclonal antibody against the serotype 4 fowl adenovirus fiber2 protein

doi:10.14088/j.cnki.issn0439-8114.2022.10.021

Abstract

Abstract: The specific monoclonal antibody against avian adenovirus type 4 （FADV-4） Fiber2 protein was prepared. The FADV-4 Fiber2 protein was purified and immunized BALB/C mice as an immunogen. Lymphocytes were isolated from spleen and fused with mouse myeloma cells SP2/0. Ten hybridoma cell lines including 1B5F1, 1B5G11, 1B5C1, 1B5D2, 1B5E3, 1B5F3, 1B5E4, 1B5G4, 1B5A8 and 1B5G8 were screened for stable secretion of monoclonal antibody against FADV-4 Fiber2 protein. Ascites were prepared from 1B5F1 cell line with the highest antibody secretion level, and the specificity was detected by Dot blot and indirect immunofluorescence assay （IFA）. The results showed that 10 hybridoma cell lines were successfully prepared. The ascites could react specifically with FADV-4 Fiber2 protein and fADV-4 challenged chicken liver tissue, which proved that the specific monoclonal antibody of FADV-4 Fiber2 protein was successfully prepared.

Key words: serotype 4 fowl adenovirus, fiber2 protein, monoclonal antibody

CLC Number:

S831.7

XIE Ming, LI Guo-pan, HU Ji-xiong, ZHANG Zhi-xiang, AI Xiao-yang, CHEN Di-si, LI Yang, YIN Jun-xiang, RONG Jun. Preparation of the monoclonal antibody against the serotype 4 fowl adenovirus fiber2 protein[J]. HUBEI AGRICULTURAL SCIENCES, 2022, 61(10): 121-126.

References

[1] NICZYPORUK J S.Phylogenetic and geographic analysis of fowl adenovirus field strains isolated from poultry in Poland[J]. Arch Virol, 2016, 161(1): 33-42.
[2] 黄国虹,许文波.腺病毒新型别的研究进展[J].病毒学报,2013, 29(3): 342-8.
[3] ZHANG T, JIN Q, DING P, et al.Molecular epidemiology of hydropericardium syndrome outbreak-associated serotype 4 fowl adenovirus isolates in central China[J]. Virol J, 2016, 13(1): 188.
[4] YE J, LIANG G, ZHANG J, et al.Outbreaks of serotype 4 fowl adenovirus with novel genotype, China[J]. Emerg Microbes Infect, 2016, 5: e50.
[5] LIU Y, WAN W, GAO D, et al.Genetic characterization of novel fowl aviadenovirus 4 isolates from outbreaks of hepatitis-hydropericardium syndrome in broiler chickens in China[J]. Emerg Microbes Infect, 2016, 5(11): e117.
[6] RUAN S, ZHAO J, HE Z, et al.Analysis of pathogenicity and immune efficacy of fowl adenovirus serotype 4 isolates[J]. Poult Sci, 2018, 97(8): 2647-2653.
[7] FEICHTNER F, SCHACHNER A, BERGER E, et al.Development of sensitive indirect enzyme-linked immunosorbent assays for specific detection of antibodies against fowl adenovirus serotypes 1 and 4 in chickens[J]. Avian Pathol, 2018, 47(1): 73-82.
[8] PEI Y, CORREDOR J C, GRIFFIN B D, et al.Fowl adenovirus 4 (FAdV-4)-based infectious clone for vaccine vector development and viral gene function studies[J]. Viruses, 2018, 10(2):97.
[9] SCHACHNER A, MAREK A, JASKULSKA B, et al.Recombinant FAdV-4 fiber-2 protein protects chickens against hepatitis-hydropericardium syndrome (HHS)[J]. Vaccine, 2014, 32(9): 1086-1092.
[10] HE Z R, RUAN S F, ZHAO J, et al.Recombinant fiber-2 protein-based indirect ELISA for antibody detection of fowl adenovirus serotype 4[J]. Avian Dis, 2018, 62(1): 73-78.
[11] WANG P, ZHANG J, WANG W, et al.A novel monoclonal antibody efficiently blocks the infection of serotype 4 fowl adenovirus by targeting fiber-2[J]. Vet Res, 2018, 49(1): 29.
[12] XIE Q, CAO S, ZHANG W, et al.A novel fiber-2-edited live attenuated vaccine candidate against the highly pathogenic serotype 4 fowl adenovirus[J]. Vet Res, 2021, 52(1): 35.
[13] YIN D, HE L, ZHU E, et al.A fowl adenovirus serotype 4 (FAdV-4) Fiber2 subunit vaccine candidate provides complete protection against challenge with virulent FAdV-4 strain in chickens[J]. Vet Microbiol, 2021, 263: 109250.
[14] 张磊,张宇,李巧.禽腺病毒4型感染的诊断与防制[J].安徽农学通报, 2016, 22(10): 116+22.
[15] 冯敬敬,牛登云,马利芳. 2015-2020年我国禽腺病毒流行病学调查及高致病性FAdV-4分离株遗传变异性分析[J].中国动物传染病学报,2021(5):92-98.
[16] 杨晶晶,金红岩.Ⅰ群禽腺病毒套式PCR检测方法的建立及应用[J].畜牧与兽医,2021, 53(9): 71-77.
[17] MOCKETT A P,COOK J K.The use of an enzyme-linked immunosorbent assay to detect IgG antibodies to serotype-specific and group-specific antigens of fowl adenovirus serotypes 2,3 and 4[J]. J Virol Methods, 1983, 7(5-6): 327-335.
[18] SHAO H, LU Y, WANG W, et al.Two novel monoclonal antibodies against fiber-1 protein of FAdV-4 and their application in detection of FAdV-4/10[J]. BMC Vet Res, 2019, 15(1): 232.
[19] 王萍. 抗血清4型禽腺病毒Fiber2蛋白单克隆抗体研制及其应用[D]. 江苏扬州:扬州大学, 2018.
[20] ZHAO J, ZHONG Q, ZHAO Y, et al.Pathogenicity and complete genome characterization of fowl adenoviruses isolated from chickens associated with inclusion body hepatitis and hydropericardium syndrome in China[J]. PLoS one, 2015, 10(7): e0133073.
[21] WANG X, TANG Q, CHU Z, et al.Immune protection efficacy of FAdV-4 surface proteins fiber-1, fiber-2, hexon and penton base[J]. Virus Res, 2018, 245: 1-6.
[22] XIA J, YAO K C, LIU Y Y, et al.Isolation and molecular characterization of prevalent Fowl adenovirus strains in southwestern China during 2015-2016 for the development of a control strategy[J]. Emerg Microbes Infect, 2017, 6(11): e103.
[23] 宋玲玲,于可响,王友令.禽腺病毒4型SYBR GreenⅠ实时荧光定量PCR检测方法的建立[J].畜牧与兽医,2019, 51(8): 118-122.
[24] ZHANG Y, LIU R, TIAN K, et al.Fiber2 and hexon genes are closely associated with the virulence of the emerging and highly pathogenic fowl adenovirus 4[J]. Emerg Microbes Infect, 2018, 7(1): 199.
[25] YU Z, AIJING L, YANAN W, et al.A single amino acid at residue 188 of hexon protein is responsible for the pathogenicity of the emerging novel fowl adenovirus 4[J]. Journal of virology, DOI:10.1128/JVI.00603-21.
[26] SHAO H, WANG P, WANG W, et al.A novel monoclonal antibodies-based sandwich ELISA for detection of serotype 4 fowl adenovirus[J]. Avian Pathol, 2019, 48(3): 204-208.
[27] 万文妍,刘延珂,王贝贝.基于FAdV-4中国流行株Fiber2蛋白的间接ELISA抗体检测方法的建立[J].中国兽医学报,2018, 38(5): 883-888.
[28] 郭瑞珍,苏冰倩,王棋.禽腺病毒血清4型Fiber2蛋白单克隆抗体制备及其抗原表位鉴定[J].畜牧兽医学报,2021,52(7): 2000-2012.
[29] GAO J, ZHAO M, DUAN X, et al.Requirement of cellular protein CCT7 for the replication of fowl adenovirus serotype 4 (FAdV-4) in leghorn male hepatocellular cells via interaction with the viral hexon protein[J]. Viruses, 2019, 11(2):107.
[30] AZIZ F, TUFAIL S, SHAH M A, et al.In silico epitope prediction and immunogenic analysis for penton base epitope-focused vaccine against hydropericardium syndrome in chicken[J]. Virus Res, 2019, 273: 197750.
[31] 刘运超,陈玉梅,冯丽丽.抗猪瘟病毒中和性单克隆抗体的制备与鉴定[J].河南农业科学,2019, 48(12): 114-120.
[32] HU J, LI G, WANG X, et al.Development of a subunit vaccine based on fiber2 and hexon against fowl adenovirus serotype 4[J]. Virus Res, 2021, 305: 198552.