Applying Microfluidics for the Production of the Cationic Liposome-Based Vaccine Adjuvant CAF09b
Subunit vaccines require particulate adjuvants to induce the desired immune responses. Pre-clinical manufacturing methods of adjuvants are often batch dependent, which complicates scale-up for large-scale good manufacturing practice (GMP) production. The cationic liposomal adjuvant CAF09b, composed...
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| Format: | Article |
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MDPI AG
2020-12-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/12/12/1237 |
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| author | Signe Tandrup Schmidt Dennis Christensen Yvonne Perrie |
| author_facet | Signe Tandrup Schmidt Dennis Christensen Yvonne Perrie |
| author_sort | Signe Tandrup Schmidt |
| collection | DOAJ |
| description | Subunit vaccines require particulate adjuvants to induce the desired immune responses. Pre-clinical manufacturing methods of adjuvants are often batch dependent, which complicates scale-up for large-scale good manufacturing practice (GMP) production. The cationic liposomal adjuvant CAF09b, composed of dioctadecyldimethylammonium bromide (DDA), monomycoloyl glycerol analogue 1 (MMG) and polyinosinic:polycytidylic acid [poly(I:C)], is currently being clinically evaluated in therapeutic cancer vaccines. Microfluidics is a promising new method for large-scale manufacturing of particle-based medicals, which is scalable from laboratory to GMP production, and a protocol for production of CAF09b by this method was therefore validated. The influence of the manufacture parameters [Ethanol] (20–40% <i>v/v</i>), [Lipid] (DDA and MMG, 6–12 mg/mL) and dimethyl sulfoxide [DMSO] (0–10% <i>v/v</i>) on the resulting particle size, colloidal stability and adsorption of poly(I:C) was evaluated in a design-of-experiments study. [Ethanol] and [DMSO] affected the resulting particle sizes, while [Lipid] and [DMSO] affected the colloidal stability. In all samples, poly(I:C) was encapsulated within the liposomes. At [Ethanol] 30% <i>v/v</i>, most formulations were stable at 21 days of manufacture with particle sizes <100 nm. An in vivo comparison in mice of the immunogenicity to the cervical cancer peptide antigen HPV-16 E7 adjuvanted with CAF09b prepared by lipid film rehydration or microfluidics showed no difference between the formulations, indicating adjuvant activity is intact. Thus, it is possible to prepare suitable formulations of CAF09b by microfluidics. |
| first_indexed | 2024-03-10T13:54:59Z |
| format | Article |
| id | doaj.art-2d06bf2c90294b5b94b1a37eda1ecb34 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-10T13:54:59Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceutics |
| spelling | doaj.art-2d06bf2c90294b5b94b1a37eda1ecb342023-11-21T01:38:33ZengMDPI AGPharmaceutics1999-49232020-12-011212123710.3390/pharmaceutics12121237Applying Microfluidics for the Production of the Cationic Liposome-Based Vaccine Adjuvant CAF09bSigne Tandrup Schmidt0Dennis Christensen1Yvonne Perrie2Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0RE, UKDepartment of Infectious Disease Immunology, Center for Vaccine Research, Statens Serum Institut, Artillerivej 5, 2300 Copenhagen S, DenmarkStrathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow G4 0RE, UKSubunit vaccines require particulate adjuvants to induce the desired immune responses. Pre-clinical manufacturing methods of adjuvants are often batch dependent, which complicates scale-up for large-scale good manufacturing practice (GMP) production. The cationic liposomal adjuvant CAF09b, composed of dioctadecyldimethylammonium bromide (DDA), monomycoloyl glycerol analogue 1 (MMG) and polyinosinic:polycytidylic acid [poly(I:C)], is currently being clinically evaluated in therapeutic cancer vaccines. Microfluidics is a promising new method for large-scale manufacturing of particle-based medicals, which is scalable from laboratory to GMP production, and a protocol for production of CAF09b by this method was therefore validated. The influence of the manufacture parameters [Ethanol] (20–40% <i>v/v</i>), [Lipid] (DDA and MMG, 6–12 mg/mL) and dimethyl sulfoxide [DMSO] (0–10% <i>v/v</i>) on the resulting particle size, colloidal stability and adsorption of poly(I:C) was evaluated in a design-of-experiments study. [Ethanol] and [DMSO] affected the resulting particle sizes, while [Lipid] and [DMSO] affected the colloidal stability. In all samples, poly(I:C) was encapsulated within the liposomes. At [Ethanol] 30% <i>v/v</i>, most formulations were stable at 21 days of manufacture with particle sizes <100 nm. An in vivo comparison in mice of the immunogenicity to the cervical cancer peptide antigen HPV-16 E7 adjuvanted with CAF09b prepared by lipid film rehydration or microfluidics showed no difference between the formulations, indicating adjuvant activity is intact. Thus, it is possible to prepare suitable formulations of CAF09b by microfluidics.https://www.mdpi.com/1999-4923/12/12/1237subunit vaccineliposomemicrofluidics |
| spellingShingle | Signe Tandrup Schmidt Dennis Christensen Yvonne Perrie Applying Microfluidics for the Production of the Cationic Liposome-Based Vaccine Adjuvant CAF09b Pharmaceutics subunit vaccine liposome microfluidics |
| title | Applying Microfluidics for the Production of the Cationic Liposome-Based Vaccine Adjuvant CAF09b |
| title_full | Applying Microfluidics for the Production of the Cationic Liposome-Based Vaccine Adjuvant CAF09b |
| title_fullStr | Applying Microfluidics for the Production of the Cationic Liposome-Based Vaccine Adjuvant CAF09b |
| title_full_unstemmed | Applying Microfluidics for the Production of the Cationic Liposome-Based Vaccine Adjuvant CAF09b |
| title_short | Applying Microfluidics for the Production of the Cationic Liposome-Based Vaccine Adjuvant CAF09b |
| title_sort | applying microfluidics for the production of the cationic liposome based vaccine adjuvant caf09b |
| topic | subunit vaccine liposome microfluidics |
| url | https://www.mdpi.com/1999-4923/12/12/1237 |
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