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Lyophilization of Protein-Loaded Polyelectrolyte Microcapsules

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ABSTRACT

Purpose

To evaluate if lyophilization can be used to obtain a dry formulation of polyelectrolyte microcapsules, which have emerged as a new class of microparticles for the encapsulation and delivery of biomacromolecules.

Methods

Microcapsules composed of dextran sulfate and poly-L-arginine were obtained by coating CaCO3 microparticles by means of the layer-by-layer technique. Microcapsules were lyophilized using different stabilizers; intactness was checked by CLSM and SEM. Horseradish peroxidase was encapsulated as model enzyme and retained activity after freeze-drying was determined using a fluorescence assay. Ovalbumin was encapsulated as model antigen; immunogenicity after lyophilization was evaluated in vitro by a T-cell proliferation assay and in vivo by measuring the antibody titer in mice.

Results

The results clearly demonstrate the necessity of using polyols in the formulation to prevent rupture of the microcapsules and to preserve the activity of encapsulated enzymes. Lyophilized microcapsules appeared as a promising adjuvant for antigen delivery, as both in vitro as in vivo assays showed higher immune activation compared to free antigen.

Conclusions

Lyophilization is a promising strategy towards improved stability of protein-loaded microcapsules.

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Abbreviations

DEXS:

dextran sulfate

HRP:

horseradish peroxidase

LbL:

layer-by-layer

OVA:

ovalbumin

pARG:

poly-L-arginine

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ACKNOWLEDGMENTS

The authors wish to thank prof Y. van Kooyk and S. Singh for performing the in vitro T-cell assays. We also wish to thank E. Adriaens for help with fluorescent assays and O. Janssens for taking SEM images. We would also like to acknowledge the Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT-Vlaanderen) for founding the BRAINSTIM project.

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Authors and Affiliations

  1. Laboratory of General Biochemistry and Physical Pharmacy Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium

    Marie-Luce De Temmerman, Joanna Rejman, Jo Demeester & Stefaan C. De Smedt

  2. Laboratory of Molecular Immunology Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium

    Johan Grooten

  3. Laboratory of Pharmaceutical Process Analytical Technology Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium

    Thomas De Beer

  4. Laboratory of Pharmaceutical Technology Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium

    Chris Vervaet

Authors
  1. Marie-Luce De Temmerman
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  2. Joanna Rejman
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  3. Johan Grooten
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  4. Thomas De Beer
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  5. Chris Vervaet
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  6. Jo Demeester
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  7. Stefaan C. De Smedt
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Correspondence to Stefaan C. De Smedt.

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De Temmerman, ML., Rejman, J., Grooten, J. et al. Lyophilization of Protein-Loaded Polyelectrolyte Microcapsules. Pharm Res 28, 1765–1773 (2011). https://doi.org/10.1007/s11095-011-0411-z

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  • DOI: https://doi.org/10.1007/s11095-011-0411-z

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