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Development of Monoclonal Antibodies and Antigen-Capture ELISA for Human Parechovirus Type 3
Keiko Goto1,2, Yutaro Yamaoka1,3, Hajera Khatun1
1Department of Microbiology, Yokohama City University School of Medicine, Kanagawa 236-0004, Japan.
Abstract:
Human parechovirus type 3 (HPeV3) is an etiologic agent of respiratory diseases, meningitis, and sepsis-like illness in both infants and adults. Monoclonal antibodies (mAbs) can be a promising diagnostic tool for antigenic diseases such as virus infection, as they offer a high specificity toward a specific viral antigen. However, to date, there is no specific mAb available for the diagnosis of HPeV3 infection. In this study, we developed and characterized mAbs specific for HPeV3 capsid protein VP0. We used cell-free, wheat germ-synthesized viral VP0 protein for immunizing BALB/c mice to generate hybridomas. From the resultant hybridoma clones, we selected nine clones producing mAbs reactive to the HPeV3-VP0 antigen, based on enzyme-linked immunosorbent assay (ELISA). Epitope mapping showed that these mAbs recognized three distinct domains in HPeV3 VP0. Six mAbs recognized HPeV3 specifically and the other three mAbs showed cross-reactivity with other HPeVs. Using the HPeV3-specific mAbs, we then developed an ELISA for viral antigen detection that could be reliably used for laboratory diagnosis of HPeV3. This ELISA system exhibited no cross-reactivity with other related viruses. Our newly developed mAbs would, thus, provide a useful set of tools for future research and ensure HPeV3-specific diagnosis.
Insights
Researchers developed specific monoclonal antibodies (mAbs) for diagnosing Human parechovirus type 3 (HPeV3) infections. This breakthrough enables reliable laboratory diagnosis of HPeV3, crucial for infant and adult respiratory and neurological illnesses.
Area of Science:
- Virology
- Immunology
- Diagnostic Development
Background:
- Human parechovirus type 3 (HPeV3) causes significant respiratory and neurological illnesses in infants and adults.
- Current diagnostic methods lack specificity for HPeV3.
- Monoclonal antibodies (mAbs) offer high specificity for viral antigen detection.
Purpose of the Study:
- To develop and characterize novel monoclonal antibodies (mAbs) specific for Human parechovirus type 3 (HPeV3).
- To establish a reliable diagnostic tool for HPeV3 infection.
Main Methods:
- Generation of hybridomas using cell-free, wheat germ-synthesized HPeV3 VP0 protein for immunization.
- Screening of hybridoma clones using enzyme-linked immunosorbent assay (ELISA).
- Epitope mapping and cross-reactivity testing of generated mAbs.
Main Results:
- Nine hybridoma clones producing HPeV3-VP0-reactive mAbs were identified.
- Six mAbs demonstrated HPeV3-specific reactivity, while three showed cross-reactivity with other parechoviruses.
- An HPeV3-specific ELISA system was developed using the generated mAbs, showing no cross-reactivity with related viruses.
Conclusions:
- Novel HPeV3-specific mAbs have been successfully developed and characterized.
- The developed mAbs are valuable tools for accurate laboratory diagnosis of HPeV3 infections.
- These mAbs will advance research and improve diagnostic specificity for HPeV3.
