Development of monoclonal antibodies targeting the conserved fragment of hexon protein to detect different serotypes of human adenovirus

Wu, Linfan1,2,3; Lin, Yuhao2,3; Yin, Juzhen4; Yang, Changbi2,3; Jiang, Yushan2,3; Zhai, Linlin2,3; Wang, Yuelin2,3; Zhu, Li2,3; Wu, Qinghua2,3; Zhang, Bao2,3; Wan, Chengsong2,3; Zhao, Wei2,3; Yang, Yang4; Shen, Chenguang2,3; Xiao, Weiwei1,2,3

2024-04-02

10.1128/spectrum.01816-23

MICROBIOLOGY SPECTRUM   影响因子和分区

2165-0497

["Human adenovirus (HAdV) infects the respiratory system, thus posing a threat to health. However, immunodiagnostic reagents for human adenovirus are limited. This study aimed to develop efficient diagnostic reagents based on monoclonal antibodies for diagnosing various human adenovirus infections. Evolutionary and homology analyses of various human adenoviral antigen genes revealed highly conserved antigenic fragments. The prokaryotic expression system was applied to recombinant penton, hexon, and IVa2 conserved fragments of adenovirus, which were injected into BALB/c mice to prepare human adenovirus-specific monoclonal antibodies. Enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), and Western blotting were used to determine the immune specificity of the monoclonal antibodies. Indirect ELISA showed that monoclonal antibodies 1F10, 8D3, 4A1, and 9B2 were specifically bound to HAdV-3 and HAdV-55 and revealed high sensitivity and low detection limits for various human adenoviruses. Western blotting showed that 1F10 and 8D3 specifically recognized various human adenovirus types, including HAdV-1, HAdV-2, HAdV-3, HAdV-4, HAdV-5, HAdV-7, HAdV-21, and HAdV-55, and 4A1 specifically recognized HAdV-1, HAdV-2, HAdV-3, HAdV-5, HAdV-7, HAdV-21, and HAdV-55. IFAs showed that 1F10, 8D3, and 4A1 exhibited highly selective localization to A549 cells infected with HAdV-3 and HAdV-55. Finally, two antibody pairs that could detect hexon antigens HAdV-3 and HAdV-55 at low concentrations were developed. The monoclonal antibodies developed in this study show potential for detecting human adenoviruses.IMPORTANCEIn this study, we selected the three most conserved antigenic fragments of human adenovirus to prepare a murine monoclonal antibody for the first time, and human adenovirus antigenic fragments with heretofore unheard of degrees of conservatism were isolated. The three monoclonal antibodies with the ability to recognize human respiratory adenovirus over a broad spectrum were screened by hybridoma and monoclonal antibody preparation. Human adenovirus infections are serious; however, therapeutic drugs and diagnostic reagents are scarce. Thus, to reduce the serious consequences of human viral infections and adenovirus pneumonitis, early diagnosis of infection is required. The present study provides three monoclonal antibodies capable of recognizing a wide range of human adenoviruses, thereby offering guidance for subsequent research and development.","In this study, we selected the three most conserved antigenic fragments of human adenovirus to prepare a murine monoclonal antibody for the first time, and human adenovirus antigenic fragments with heretofore unheard of degrees of conservatism were isolated. The three monoclonal antibodies with the ability to recognize human respiratory adenovirus over a broad spectrum were screened by hybridoma and monoclonal antibody preparation. Human adenovirus infections are serious; however, therapeutic drugs and diagnostic reagents are scarce. Thus, to reduce the serious consequences of human viral infections and adenovirus pneumonitis, early diagnosis of infection is required. The present study provides three monoclonal antibodies capable of recognizing a wide range of human adenoviruses, thereby offering guidance for subsequent research and development."]

human adenovirus hexon protein monoclonal antibodies ELISA Western blot indirect immunofluorescence assay

SCI

英语

通讯

MOST | National Natural Science Foundation of China (NSFC)["32170939","82371846"] ; National Natural Science Foundation of China[2022B1515020075] ; Guangdong Basic and Applied Basic Research Foundation[2021B1212030014] ; Guangdong Science and Technology Program key projects[202201011419]

Microbiology

Microbiology

WOS:001167430100001

AMER SOC MICROBIOLOGY

Web of Science

1.Guangdong Med Univ, Sch Publ Hlth, Zhanjiang, Peoples R China
2.Southern Med Univ, Sch Publ Hlth, BSL Lab Guangdong 3, Guangdong Prov Key Lab Trop Dis Res, Guangzhou, Peoples R China
3.Southern Med Univ, Zhujiang Hosp, Dept Lab Med, Guangzhou, Peoples R China
4.Southern Univ Sci & Technol, Shenzhen Peoples Hosp 3, Shenzhen Key Lab Pathogen & Immun, State Key Discipline Infect Dis,Hosp 2, Shenzhen, Peoples R China

Wu, Linfan,Lin, Yuhao,Yin, Juzhen,et al. Development of monoclonal antibodies targeting the conserved fragment of hexon protein to detect different serotypes of human adenovirus[J]. MICROBIOLOGY SPECTRUM,2024,12(4).

Wu, Linfan.,Lin, Yuhao.,Yin, Juzhen.,Yang, Changbi.,Jiang, Yushan.,...&Xiao, Weiwei.(2024).Development of monoclonal antibodies targeting the conserved fragment of hexon protein to detect different serotypes of human adenovirus.MICROBIOLOGY SPECTRUM,12(4).

Wu, Linfan,et al."Development of monoclonal antibodies targeting the conserved fragment of hexon protein to detect different serotypes of human adenovirus".MICROBIOLOGY SPECTRUM 12.4(2024).