PROJECT

Plant-produced subviral HBV particles as carriers for antigen protein epitopes for the

purpose of vaccines with increased immunogenicity.

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

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

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.

Research progress

Preparation of vectors forAgrobacterium tumefaciens

Purification of antigens for injection

GMO of tobaccoproducing HBc-preS2 antigen

Generativeand/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-preS2868pz

868pz

1400pz

1000pz750pz500pz

300pz

2000pz

1400pz

1000pz750pz500pz300pz

2000pz

3000pz4000pz6000pz

10000pz

Ścieżki:M – DNA size marker5 etc.

HBs – positive control

- trangenic plants L-N – non-transgenic plantsW – negative control

Transgenic plants PCR analysisHBcAg-preS2 content

in transgenic plants