Freeze-drying cycle optimization for the rapid preservation of protein-loaded liposomal formulations

Maryam T Hussain, Neil Forbes, Yvonne Perrie, Kiran P Malik, Chinwe Duru, Paul Matejtschuk

Research output: Contribution to journalArticle

Abstract

Technology such as the use of microfluidics to generate liposomes has been well researched, yet the stabilisation of liposomal formulations is a major challenge to their greater implementation. To the best of our knowledge, this is the first study investigating the use of 96 well plates to freeze-dry ovalbumin (OVA) loaded neutral (DMPC:Chol and DSPC:Chol), anionic (DSPC:Chol:PS) and cationic (DSPC:Chol:DOTAP) liposomes. Through the use of high throughput screening, a freeze drying cycle was optimised; ramp freezing from from 4°C to -45°C, followed by primary drying at -30°C and secondary drying at 30°C under a vacuum of 0.1 mBar. These parameters maintained liposome physicochemical properties, with the liposomes remaining below 100 nm and were homogenous (polydispersity index of less than 0.2 post rehydration). Minimal leakage of the OVA protein was observed, with almost 100% OVA remaining encapsulated post rehydration of the formulations. Here we have identified a simple method that allows for the rapid screening and freeze-drying of a range of liposomal formulations.
LanguageEnglish
JournalInternational Journal of Pharmaceutics
Publication statusAccepted/In press - 19 Sep 2019

Fingerprint

Freeze Drying
Liposomes
Ovalbumin
Fluid Therapy
Proteins
Dimyristoylphosphatidylcholine
Architectural Accessibility
Microfluidics
Vacuum
Freezing
Technology

Keywords

  • liposomes
  • stabilisation
  • freeze-dry
  • OVA protein

Cite this

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title = "Freeze-drying cycle optimization for the rapid preservation of protein-loaded liposomal formulations",
abstract = "Technology such as the use of microfluidics to generate liposomes has been well researched, yet the stabilisation of liposomal formulations is a major challenge to their greater implementation. To the best of our knowledge, this is the first study investigating the use of 96 well plates to freeze-dry ovalbumin (OVA) loaded neutral (DMPC:Chol and DSPC:Chol), anionic (DSPC:Chol:PS) and cationic (DSPC:Chol:DOTAP) liposomes. Through the use of high throughput screening, a freeze drying cycle was optimised; ramp freezing from from 4°C to -45°C, followed by primary drying at -30°C and secondary drying at 30°C under a vacuum of 0.1 mBar. These parameters maintained liposome physicochemical properties, with the liposomes remaining below 100 nm and were homogenous (polydispersity index of less than 0.2 post rehydration). Minimal leakage of the OVA protein was observed, with almost 100{\%} OVA remaining encapsulated post rehydration of the formulations. Here we have identified a simple method that allows for the rapid screening and freeze-drying of a range of liposomal formulations.",
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AU - Forbes, Neil

AU - Perrie, Yvonne

AU - Malik, Kiran P

AU - Duru, Chinwe

AU - Matejtschuk, Paul

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AB - Technology such as the use of microfluidics to generate liposomes has been well researched, yet the stabilisation of liposomal formulations is a major challenge to their greater implementation. To the best of our knowledge, this is the first study investigating the use of 96 well plates to freeze-dry ovalbumin (OVA) loaded neutral (DMPC:Chol and DSPC:Chol), anionic (DSPC:Chol:PS) and cationic (DSPC:Chol:DOTAP) liposomes. Through the use of high throughput screening, a freeze drying cycle was optimised; ramp freezing from from 4°C to -45°C, followed by primary drying at -30°C and secondary drying at 30°C under a vacuum of 0.1 mBar. These parameters maintained liposome physicochemical properties, with the liposomes remaining below 100 nm and were homogenous (polydispersity index of less than 0.2 post rehydration). Minimal leakage of the OVA protein was observed, with almost 100% OVA remaining encapsulated post rehydration of the formulations. Here we have identified a simple method that allows for the rapid screening and freeze-drying of a range of liposomal formulations.

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