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Use of Viral Entry Assays and Molecular Docking Analysis for the Identification of Antiviral Candidates against Coxsackievirus A16
Published on: July 15, 2019
A uniform quantitative enzyme-linked immunosorbent assay for Coxsackievirus A16 antigen in vaccine
Bopei Cui1, Fang Cai2, Fan Gao1
1Divison of Hepatitis Virus and Enterovirus Vaccine, National Institutes for Food and Drug Control, Institute for Biological Product Control , Beijing, China.
Insights
A new quantitative enzyme-linked immunosorbent assay (Q-ELISA) was developed to measure Coxsackievirus A16 (CV-A16) antigen concentration in vaccines. This method is crucial for the quality control of hand, foot, and mouth disease (HFMD) vaccines.
Area of Science:
- Virology
- Immunology
- Vaccine Development
Background:
- Coxsackievirus A16 (CV-A16) is a primary cause of hand, foot, and mouth disease (HFMD) outbreaks globally, particularly in children.
- Effective prevention and control of HFMD rely on vaccine development, but quality control is hindered by the absence of a standardized CV-A16 antigen detection method.
Purpose of the Study:
- To establish and validate a quantitative enzyme-linked immunosorbent assay (Q-ELISA) for determining CV-A16 antigen concentration.
- To enable reliable quality control (QC) for CV-A16 vaccines manufactured in China.
Main Methods:
- Development of a Q-ELISA using a neutralizing antibody (16E1) as the capture antibody and a HRP-conjugated antiserum for detection.
- Validation of the Q-ELISA for specificity, linearity, accuracy, precision, and robustness using the CV-A16 antigen national standard (NS).
- Application of the Q-ELISA to quantify antigen content in vaccine bulks from six manufacturers and intermediate products.
Main Results:
- The developed Q-ELISA demonstrated validated performance characteristics (specificity, linearity, accuracy, precision, robustness).
- The assay successfully quantified CV-A16 antigen levels in vaccine samples from multiple manufacturers.
- Results confirmed the Q-ELISA's suitability for meeting the QC requirements of involved manufacturers.
Conclusions:
- The established Q-ELISA provides a reliable method for CV-A16 antigen quantification.
- This assay is essential for ensuring the quality and effectiveness of CV-A16 vaccines.
- The Q-ELISA supports the manufacturing and quality control of HFMD vaccines in China.
Abstract:
Coxsackievirus A16 (CV-A16), one of major etiological agents of hand, foot and mouth disease (HFMD), causes outbreaks of the disease in young children all over the world. In order to promote the prevention and control of HFMD, the research and development of CV-A16 vaccine have been carried out in China. However, due to lacking of a recognized CV-A16 antigen detection method, the evaluation and quality control (QC) of vaccine effectiveness are greatly limited. In this study, we established a quantitative enzyme-linked immunosorbent assay (Q-ELISA) to determine the antigen concentration in CV-A16 vaccines that can be applied in manufacturing in China. A neutralizing antibody 16E1 was used as a capture antibody that can bind to various CV-A16 antigens of different subgenotypes, and an antiserum from CV-A16-immunized rabbit conjugated by HRP was suitable for detecting and quantifying CV-A16 antigens. The Q-ELISA was validated for specificity, linearity, accuracy, precision and robustness by using the CV-A16 antigen national standard (NS). Furthermore, we utilized the Q-ELISA to quantify antigen contents of vaccine bulks from six manufacturers and other intermediate products from one manufacturer. The results indicated that the Q-ELISA can satisfy the requirements of QC for all manufacturers involved.
