AFA-Liposomes(TM)

Adaptive Focused Acoustics™ (AFA) has been shown to be effective in the formation of mono-disperse liposomes. Liposomes are being used to deliver a number of different active pharmaceuticals for both small and large molecules.

Traditional liposome preparation methods include detergent depletion, ethanol injection, reverse-phase evaporation and emulsification. Processing methods include high-pressure homogenization, extrusion and ultrasound. The major disadvantages of these preparation methods include the consumption of large volumes of volatile organic solvents, multiple lengthy steps and potential degradation of the samples due to heat exposure.

AFA offers a breakthrough alternative over these traditional methods. AFA has demonstrated the ability to form monodisperse liposomes at 4°C (below the lipid phase transition temperature) and also without the use of organic solvents.

AFA can be used to form mono-disperse liposomes using a range of lipids and formulations, including the addition cholesterol and other difficult to load materials. Active ingredients can be loaded into the liposome at the time of formation, allowing a very simple, and effective, one step Formulation within a matter of minutes, without the risk of cross contamination or the need to cleanup.

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[tab title=”Features
& Benefits”]

Co-solvent Free
No need to dissolve lipids or hydrophobic compounds in a co-solvent. All the materials can be included in the aqueous buffer, and then formulated.

Low-Temperature Formulation
Liposomes can be formed at 4C, irrespective of the lipid phase transition temperature.

Delicate Compounds
Energy levels can be pre-set to enable formulation of delicate macromolecules such as siRNA and proteins.

Short Processing Time
One-step formulation using an automated apparatus.

No Cross Contamination
Liposomes are produced in single-use autoclavable glass vials.

Automated Size Control (under development)
Real-time monitoring and feedback control of particle size.

Scalable
Capable of scaling from 100 microliters through continuous flow processing.

 

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[tab title=”Complementary
Products”]

Covaris S-Series benchtop system
Single vessel processing from 300ul through 18ml sample volumes:
• Discovery
• Animal dosing
• Method development
Covaris FS220-M
Rack System
Rack System
Automated processing in a comprehensive system design
• Continuous Flow Cell System
• Thermal and dissovled oxygen monitoring
• Flow rates up to 250mL/minute
Covaris FS-Series Benchtop laboratory system
Processing single vessels up to 18ml. Processing batch material from 22ml through multi-liter
• Discovery to Development
• Larger animal study dosing
• Process method development
Covaris FS220-C
Pilot scale system with data acquisition monitoring of continuous process streams.
Processing single vessels up to 18ml. Processing batch material from 22ml through multi-liter
• Discovery to Development
• Larger animal study dosing to First Human trial
• Process method development
• Continuous process with PAT analytical feedback

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[tab title=”AFA-Liposome
Protocols”]

DOPE Based AFA Liposome

HEL Based AFA Liposome

Phospholipon 90G Based AFA Liposome

Blank Doxil formulation AFA Liposome

Phospholipon 90G Based AFA Lipsome with 26 nanometer average particle size

 

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[tab title=”References
…”]

Making unilamellar liposomes using focused ultrasound. Tejera-Garcia R etal. Langmuir. 2011 Aug 16;27(16):10088-97. Epub 2011 Jul 21.

In this paper Scientist from Aalto University Finland used Covaris AFA to form liposomes and they reconfirmed that the Covaris technology “provides a simple and easy to use approach for making liposomes with several advantages: it is minimally invasive and involves no loss of material. Precisely controlled wavelengths are employed with a significant reduction in the presence of hot spots, which could destroy some biological materials of interest.

http://www.ncbi.nlm.nih.gov/pubmed?term=Making%20Unilamellar%20Liposomes%20Using%20Focused%20Ultrasound

Oxidized phosphatidylcholines facilitate phospholipid flip-flop in liposomes. Volinsky R etal. Biophys J. 2011 Sep 21; 101(6):1376-84. Epub 2011 Sep 20.

In this paper authors mention that they used Covaris technology for making Single uni lamellar vesicles.

http://www.ncbi.nlm.nih.gov/pubmed?term=Oxidized%20Phosphatidylcholines%20Facilitate%20Phospholipid%20Flip-Flop%20in%20Liposomes

Peptide-mediated targeting of liposomes to TrkB receptor-expressing cells. Sanjeev Ranjan etal. Int J Nanomedicine. 2012; 7: 3475–3485.

In this paper authors used Covaris technology in the “Incorporation of peptide-PEG-lipid conjugates into liposomes” after the organic solvent evaporation step.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3405886/

Preclinical Nanomedicine: Internalization of liposome nanoparticles functionalized with TrkB ligand in rat cochlear cell populations. Jing Zou etal. European Journal of Nanomedicine. Volume 2, Issue 2, Pages 7–13.

In this paper Scientist used Covaris technology to prepare liposomal nanoparticle.

http://www.degruyter.com/dg/viewarticle/j$002fejnm.2009.2.2$002fejnm.2009.2.2.7$002fejnm.2009.2.2.7.xml;jsessionid=D9770769A689FCC1115430614ED07109

Liposome Preparation Technology:
Freeze Fracture Electron Microscopic Examination & Analysis of Liposomes Produced by Covaris AFA Tecnology 

Sensitivity of nanostructure in charged cubosomes to phase changes triggered by ionic species in solution. Quingtao Liu et al. Langmuir 10 Oct 2013; DOI: 10.1021/la402426y
http://pubs.acs.org/doi/pdf/10.1021/la402426y

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[tab title=”Case
Studies”]

Liposome Formation Using AFA Technology

Manufacturing Liposomes from Various Lipid Blends Using AFA Technology

DPPC: Cholesterol Liposome Formulation

Freeze Fracture Electron Microscopy of Liposomes Using AFA Technology

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