Simultaneous In Situ Visualization and Quantitation of Dual Antigens Adsorbed on Adjuvants Using High Content Analysis

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.

Keywords:

• adjuvant
• in situ antigenicity
• in vitro relative potency
• monoclonal antibody
• multiplexed fluorescence immunoassay
• virus-like particles

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at:www.tandfonline.com/doi/full/10.2217/epi-2016-0184

Author contributions

Q Zhao and N Xia designed the study. Z Zhang, L Cao, X Wang, J Cao, X Huang, Y Cai, Z Lin, H Pan and M Fang performed the experiments. Z Zhang, T Zhang, L Cao and Q Zhao analyzed the data. Z Zhang, T Zhang and L Cao wrote the manuscript. Z Zhang, T Zhang, L Cao, Q Yuan, S Li, J Zhang, N Xia and Q Zhao participated in discussion and interpretation of the results. All authors read and approved the final manuscript.

Acknowledgments

The authors would like to thank to Ms Xiaoling Li for her invaluable technical support in high content imaging analysis and flow cytometry analysis.

Financial & competing interests disclosure

This work was supported by the National Natural Science Foundation of China (grant numbers 31670939, 31730029 and 31670935) and the Scientific Research Foundation of the State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics (grant number 2018ZY001). H Pan and Z Lin are the Xiamen Innovax Biotech Company employees. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

This study was carried out the approval by the Xiamen University Animal Care and Use Committee. The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations.