Simultaneous in situ visualization and quantitation of dual antigens adsorbed on adjuvants using high content
Zhigang Zhang1, Tianying Zhang1, Lu Cao1
1State Key Laboratory of Molecular Vaccinology & Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, Fujian 361105, PR China.
Insights
A new in situ method analyzes vaccine antigenicity directly on adjuvants, simplifying analysis. This approach allows for simultaneous visualization and quantification of antigens, improving vaccine development.
Area of Science:
- Vaccinology
- Immunology
- Biotechnology
Background:
- Traditional antigenicity assays require antigen recovery from particulate adjuvants.
- This limits the efficiency and accuracy of vaccine analysis.
Purpose of the Study:
- To develop an in situ method for interrogating vaccine antigens adsorbed on adjuvants.
- To enable direct analysis of antigenicity without prior antigen recovery.
Main Methods:
- Utilized fluorescence imaging-based high-content analysis.
- Visualized antigen distribution on adjuvant agglomerates.
- Analyzed antigenicity of adsorbed antigens using monoclonal antibodies.
Main Results:
- Achieved simultaneous visualization and quantification of dual antigens in a bivalent human papillomavirus vaccine.
- Demonstrated good agreement between in situ multiplexed assays and traditional sandwich ELISA.
- Successfully analyzed antigenicity of adsorbed antigens.
Conclusions:
- The in situ antigenicity analysis offers streamlined procedures for vaccine assessment.
- Its amenability for multiplexing makes it a favorable choice for in vitro functional assessment.
- This method is suitable for bionanoparticles used as vaccine antigens.
Abstract:
Aim: Traditional antigenicity assay requires antigen recovery from the particulate adjuvants prior to analysis. An in situ method was developed for interrogating vaccine antigens with monoclonal antibodies while being adsorbed on adjuvants. Materials & methods: The fluorescence imaging-based high content analysis was used to visualize the antigen distribution on adjuvant agglomerates and to analyze the antigenicity for adsorbed antigens. Results: Simultaneous visualization and quantitation were achieved for dual antigens in a bivalent human papillomavirus vaccine with uniquely labeled antibodies. Good agreement was observed between the in situ multiplexed assays with well-established sandwich enzyme-linked immunosorbent assays. Conclusion: The streamlined procedures and the amenability for multiplexing make the in situ antigenicity analysis a favorable choice for in vitro functional assessment of bionanoparticles as vaccine antigens.
