Presentation of a scientific research project - expression of a fusion proteins in plants - carrier structure for immunogenic epitopes, for the purpose of oral vaccine, but also for potential purification and traditional vaccine.

1. PROJECT
Plant-produced subviral HBV particles as
carriers for antigen protein epitopes for the
purpose of vaccines with increased
immunogenicity.

2. Worldwide need for a therapeutic vaccine against chronic HBV
infections as THERE ARE OVER 370 MLN OF CHRONIC SICK
worldwide who often develop liver dysfunction, cirrhosis and
cancer.
Shepard CW et al. (2006)
HBV infected population 2-8 %
< 2 %
> 8 %
Protein production in plants:
• Low cost of production – comparable with microbiological production
• Activity - proteins expressed in native and biologically active forms
• Safety – plants cannot transmit human pathogens

3. HBcAg Capsid-Like Particles
HBcAg as a carrier for immunogenic epitopes
Acc. James A. Perkins, 2002
• Plant–expressed HBcAg forms Capsid-Like Particles, very durable and
immunogenic structures and a good platform for presenting immunogenic
epitopes
• PreS2 domain - structural element of surface M- and L-HBs antigens, which
induces high immunological response in 99,5% of population
PreS2 domain
Based on Wynne, 1999
HBcAg dimer
PreS2 domain

4. Research objectives
 Transformation of Nicotiana tabacum to obtain
genetically modified plants expressing HBcAg–preS2
protein
 Analysis of HBcAg–preS2 expression level and form of
obtained transformed plants.
 Selection of highest expressing plants for material
propagation and further tests.

5. Research progress
Preparation of vectors for
Agrobacterium tumefaciens
Purification of antigens
for injection
GMO of tobacco
producing HBc-preS2 antigen
Generative
and/or vegetative
propagationPlant
transformation
and regeneration
plazmid Ti
wektor binarny
T-DNA
chromosom
Plants analysis
0
100
200
300
4J/3 7H/712B/1013G/33
[mg/g FW of leaves]
HBcAg-preS2
868pz
868pz
1400pz
1000pz
750pz
500pz
300pz
2000pz
1400pz
1000pz
750pz
500pz
300pz
2000pz
3000pz
4000pz
6000pz
10000pz
Ścieżki:
M – DNA size marker
5 etc.
HBs – positive control
- trangenic plants
L-N – non-transgenic plants
W – negative control
Transgenic plants PCR analysis
HBcAg-preS2 content
in transgenic plants