Translating a Thin-Film Rehydration Method to Microfluidics for the Preparation of a SARS-CoV-2 DNA Vaccine: When Manufacturing Method Matters
Allegra Peletta,
Eakachai Prompetchara,
Kittipan Tharakhet,
Papatsara Kaewpang,
Supranee Buranapraditkun,
Nongnaphat Yostrerat,
Suwimon Manopwisedjaroen,
Arunee Thitithanyanont,
Jonathan Avaro,
Leonard Krupnik,
Antonia Neels,
Kiat Ruxrungtham,
Chutitorn Ketloy,
Gerrit Borchard
Affiliations
Allegra Peletta
Section of Pharmaceutical Sciences, Institute of Pharmaceutical Sciences of Western Switzerland (ISPSO), University of Geneva, 1211 Geneva, Switzerland
Eakachai Prompetchara
Department of Laboratory Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
Kittipan Tharakhet
Department of Laboratory Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
Papatsara Kaewpang
Center of Excellence in Vaccine Research and Development (Chula VRC), Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
Supranee Buranapraditkun
Center of Excellence in Vaccine Research and Development (Chula VRC), Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
Nongnaphat Yostrerat
Center of Excellence in Vaccine Research and Development (Chula VRC), Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
Suwimon Manopwisedjaroen
Department of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand
Arunee Thitithanyanont
Department of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand
Jonathan Avaro
Empa, Swiss Federal Laboratories for Materials Science and Technology, Center for X-ray Analytics, Lerchenfeldstrasse 5, 9014 St. Gallen, Switzerland
Leonard Krupnik
Empa, Swiss Federal Laboratories for Materials Science and Technology, Center for X-ray Analytics, Lerchenfeldstrasse 5, 9014 St. Gallen, Switzerland
Antonia Neels
Empa, Swiss Federal Laboratories for Materials Science and Technology, Center for X-ray Analytics, Lerchenfeldstrasse 5, 9014 St. Gallen, Switzerland
Kiat Ruxrungtham
Center of Excellence in Vaccine Research and Development (Chula VRC), Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
Chutitorn Ketloy
Department of Laboratory Medicine, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
Gerrit Borchard
Section of Pharmaceutical Sciences, Institute of Pharmaceutical Sciences of Western Switzerland (ISPSO), University of Geneva, 1211 Geneva, Switzerland
Previous investigations conducted on a liposomal formulation for a SARS-CoV-2 DNA vaccine manufactured using the thin-film layer rehydration method showed promising immunogenicity results in mice. The adaptation of the liposomal formulation to a scalable and reproducible method of manufacture is necessary to continue the investigation of this vaccine candidate. Microfluidics manufacture shows high potential in method translation. The physicochemical characterization of the blank liposomes produced by thin-film layer rehydration or microfluidics were shown to be comparable. However, a difference in lipid nanostructure in the bilayer resulted in a significant difference in the hydration of the thin-film liposomes, ultimately altering their complexation behavior. A study on the complexation of liposomes with the DNA vaccine at various N/P ratios showed different sizes and Zeta-potential values between the two formulations. This difference in the complexation behavior resulted in distinct immunogenicity profiles in mice. The thin-film layer rehydration-manufactured liposomes induced a significantly higher response compared to the microfluidics-manufactured samples. The nanostructural analysis of the two samples revealed the critical importance of understanding the differences between the two formulations that resulted in the different immunogenicity in mice.
