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Epitope mapping and characterization of a neutralizing monoclonal antibody against human adenovirus type 3
Xingui Tian1, Chenyang Li, Chunyan Xue
1State Key Lab of Respiratory Disease, Guangzhou Institute of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510120, China.
Abstract:
Human adenovirus serotype 3 (HAdV-3) has occurred as a global epidemic in recent years causing serious diseases such as pneumonia in pediatric and adult patients. Development of reliable diagnostic reagents and identification of neutralizing epitopes is important for the surveillance and control of infection. In this study, a neutralizing monoclonal antibody (MAb) MAb 1B6 was generated using the HAdV-3 virion. MAb 1B6 specially recognized the HAdV-3 virus particles and the HAdV-3 hexon protein, but not the virus particles or the hexon protein of HAdV-7 and HAdV-4 by western-blot analysis and indirect enzyme-linked immunosorbent assay (ELISA). Analysis using a series of peptides from the hexon protein and chimeric adenovirus (Ad) particles of epitope mutants revealed that MAb 1B6 bound to the exposed region (amino acid positions 414-424 of hexon) in hypervariable region 7 (HVR7). ELISA demonstrated that MAb 1B6 could recognize the corresponding regions of other HAdV-3 genotypes that have some residues substituted. The identification of the neutralizing epitope and the generation of MAb 1B6 may be useful for clinical serotype-specific diagnosis, subunit vaccine construction for HAdV-3 infection, and virion structural analysis.
Insights
Researchers developed a neutralizing monoclonal antibody (MAb) targeting Human adenovirus serotype 3 (HAdV-3). This MAb identifies a specific epitope on the HAdV-3 hexon protein, aiding in diagnostics and vaccine development for HAdV-3 infections.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Human adenovirus serotype 3 (HAdV-3) causes global epidemics, leading to severe pneumonia in various age groups.
- Effective control requires reliable diagnostic tools and identification of neutralizing epitopes for HAdV-3 surveillance.
- Monoclonal antibodies (MAbs) are crucial for precise viral detection and characterization.
Purpose of the Study:
- To generate a neutralizing monoclonal antibody (MAb) against HAdV-3.
- To identify the specific neutralizing epitope on the HAdV-3 virion.
- To evaluate the MAb's utility in diagnostics and potential vaccine development.
Main Methods:
- Generation of a neutralizing MAb (MAb 1B6) using HAdV-3 virions.
- Western-blot and indirect ELISA to assess MAb specificity against HAdV-3 and related serotypes (HAdV-7, HAdV-4).
- Epitope mapping using hexon protein peptides and chimeric adenovirus particles to pinpoint the binding site.
Main Results:
- MAb 1B6 specifically recognized HAdV-3 virus particles and the HAdV-3 hexon protein, showing no cross-reactivity with HAdV-7 or HAdV-4.
- The MAb bound to an exposed region (amino acids 414-424) within hypervariable region 7 (HVR7) of the hexon protein.
- MAb 1B6 successfully recognized corresponding regions in other HAdV-3 genotypes with minor residue substitutions.
Conclusions:
- A neutralizing MAb, MAb 1B6, was successfully generated against HAdV-3.
- The study identified a specific neutralizing epitope on the HAdV-3 hexon protein (HVR7).
- This MAb and epitope information are valuable for HAdV-3 serotype-specific diagnostics, subunit vaccine design, and structural analysis.
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