Presentation on theme: "Potato Vaccine against Hepatitis B PLANT & MAMMALIAN CELL TECHNOLOGY BSB3163 13 TH DEC 2011 BY: KALAISELVI MOHANRAJSB09031 RUBINI DEVI SELVARAJOOSB09005."— Presentation transcript:
Potato Vaccine against Hepatitis B PLANT & MAMMALIAN CELL TECHNOLOGY BSB3163 13 TH DEC 2011 BY: KALAISELVI MOHANRAJSB09031 RUBINI DEVI SELVARAJOOSB09005 SUMITHALAKSMY GUNASEGARANSB09063
BACKGROUND Hepatitis B is a viral infection that attacks the liver and can cause both acute and chronic disease. The virus is transmitted through contact with the blood or other body fluids of an infected person - not through casual contact. (Fact sheet N°204 Revised August 2008)
Over two billion people worldwide are infected with hepatitis B, a serious liver infection that can result in Jaundice Cirrhosis and Liver cancer. (ISB News Report, February 2001)
The current hepatitis vaccine extracted from yeast (injectable vaccine) requires chemical modification to become active, increasing the cost of the vaccine, which also must be stored under refrigeration. This has severely limited its utility to more than one-third of the world's population, especially in third world regions where the disease in rampant. The situation is further complicated by the need for three separate injections of the vaccine at 0, 1, and 6 months of age. (ISB News Report, February 2001)
Dr. Charles Arntzen, of Arizona State University, has generated and forward the idea of a stable, plant-based vaccine as an attractive alternative. Dr. Charles Arntzen and his colleagues studied various ways to increase plant production of the Hepatitis B antigen, HbsAg, in potato. (ISB News Report, February 2001)
The transgenic potatoes were created and grown by Dr. Charles J. Arntzen and Hugh S. Mason and their colleagues at the Boyce Thompson Institute for Plant Research, an affiliate of Cornell University, USA. (ISB News Report, February 2001)
Dr. Charles J. Arntzen and his colleagues have took a gene out of the hepatitis B virus and incorporated it in the potato plant, which responded by producing the Hepatitis B virus antigen. Once ingested, this antigen protein creates an immune response in the human body that acts as a booster shot against the Hepatitis B virus. (Science News, 2000)
Dominated clinical trials Easily manipulated/transformed Easily propagated from its eyes Stored for long periods without refrigeration Easy for mass production system by breeding compared to an animal system Possible production of vaccines with low costs Reduced need for medical personnel and sterile injection conditions Economical to mass produce and transport Heat stable, eliminating the need for refrigeration Antigen protection through bioencapsulation Subunit vaccine (not attenuated pathogens) means improved safety
Dosage of vaccines would be variable. Not convenient for infants. Needs cooking which can denature antigen and decrease immunogenicity *Some kinds of South American potatoes can be eaten raw. Although some studies show that cooking does not destroy full complement of antigen in potatoes.
Edible vaccines are thought to be possible and promising yet there are still some issues!
Long term effect of those who consume transgenic plants are not known Some say playing God by making organisms produce things they normally would not produce is unnatural and wrong Ethical Issues
Safety Issues Plant/crops (food) contamination through cross pollination and of vaccine itself in plant debris spreading dust and other pollutant in surfaces and ground waters. The vaccine antigen may affect browsing animals and humans living in the area drinking vaccine polluted water or breathing vaccine polluted dust.
The cultivation and production of pharmaceutical crops should be limited to control the production facilities like greenhouse, or in plant tissue culture, that prevent the environmental release of biopharmaceuticals
a dosage problem might occur, as the production of antigens is likely to vary from plant to plant. A possible way to overcome this would involve processing the transgenic plants into concentrated forms, so that dosing could be uniform. More studies need to be performed in order to determine the safety of edible vaccines, especially since horizontal gene transfer may occur, thus increasing the risk of creating new strains of viruses.
COMPANY Wisconsin by American Ag- Tec International, Ltd., a Delavan, Wisconsin based pharmaceutical agricultural technology company in NY. Axis Genetics Cambridge, England, a plant pharmaceutical company now holds an exclusive License for this medical technology. Boyce Thompson Institute for Plant Research in Ithaca, New York develop Edible Plant Vaccines for the prevention of Hepatitis B. (SeedQuest 13 th Nov 1998)
Distilled water FeSO 4.7H 2 O Na 2 EDTA NaOH MS macronutrient MS micronutrient Gamborgs B5 vitamin Sucrose Agar Buffer solutions (pH 4, pH 7 and pH 10) 2,4-Dichlorophenoxyacetic acid (2,4-D) Benzo(a)pyrene (BAP) NH 4 NO 3 CaCl 2.2H 2 O MgSO 4.7H 2 O KH 2 PO 4 H 3 BO 3 MnSO 4.H 2 O ZnSO 4.7H 2 O KI Na 2 MoO 4.2H 2 O CuSO 4.5H 2 O CoCl 2.6H 2 O Glycine Myo-inositol Nicotinic acid Pyridoxine Thiamine HCl 0.1 M NaOH Commercial ethanol
Beakers 250 mL Beakers 500 mL Electronic Balance Spatula Magnetic stirrer Measuring cylinder, 100 mL Measuring cylinder, 500 mL Measuring cylinder, 1000 mL Microwave Sterile jars (X10) Autoclave Reagent bottles (1000 mL and 100 mL) Blue cap Scott bottle 500 mL Reagent bottles Pipettes Refrigerator Volumetric Flask Culture plates Potato explant Knife Forceps Bunsen Burner Aluminium Foil Para film tape
METHOD(S) Extract virus strain from HBV and inactivate it into HBsAg Clone HBsAg into pROK2S shuttle plasmid Electroporate the plasmid into Agrobacterium tumefaciens Plate the bacterium and make colonies
Co-cultivate the leaf fragment to induce callus using gel medium Transfer explant grown into soil Select a potato explant and introduce the Agrobacterium into the explant
REFERENCES Kapusta.J, Modelska. A, et.al, A Plant Derived Edible Vaccine Against Hepatitis B Virus, The Faseb Journal, 13: 1796-1798, Oct 1999. Kirk D.D, McIntosh.K, Walmsley A, et.al, Risk Analysis for Plant Made Vaccines, Springer Transgenic Research, 14:449–462, 2005. ISB News Report, February 2001(online) http://www.biotechinfo.net/hepatitus_vaccine.html American Ag-Tec International Ltd, 2006(online) http://www.ag-tec.com/potato.htm