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BioMed Research International
Volume 2014, Article ID 485689, 10 pages
Research Article

Freeze-Drying of Plant Tissue Containing HBV Surface Antigen for the Oral Vaccine against Hepatitis B

1Institute of Plant Genetics, Polish Academy of Sciences, Strzeszyńska 34, 60-479 Poznań, Poland
2Poznań University of Life Sciences, Wojska Polskiego 28, 60-995 Poznań, Poland
3Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Rudolfa Weigla 12, 53-114 Wrocław, Poland

Received 16 May 2014; Revised 11 September 2014; Accepted 17 September 2014; Published 12 October 2014

Academic Editor: Héla Kallel

Copyright © 2014 Marcin Czyż et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

To assess excipient potency for stabilisation of S-HBsAg during the freeze-drying process series of substances were tested with regard to their reported lyoprotective properties and requirements of oral vaccine formulation. For the preliminary test, fresh lettuce leaves were infiltrated with sucrose, glucose, mannitol, glycine and glycerol at concentrations of 100, 250 and 500 mM under 100 mbar vacuum for 5 min and lyophilised at 5°C and 20°C for primary drying and 22°C for secondary drying.

Basing on absolute antigen values and relative preservation, it could be stated that across all tested variants sucrose and then mannitol at highest concentrations appeared as the most suitable for lyoprotection of S-HBsAg. Statistical analysis revealed that sucrose 500 mM was the most effective variant regarding relative content of VLPs in both drying profiles. As a consequence this protectant was chosen for further testing as expressing the best potential of S-HBsAg preservation, even when used in the unfavourable drying profile.

  1. Supplementary Table