Molecular farming

LIBARDO CARABALLO BLANCO

BIOTECNOLOGIA I 2009

MOLECULAR FARMING

Biopharming / Molecular Farming / Gene Farming

• Production of biopharmaceutical and biologic products for human and animal health in transgenic plants

• The products of molecular farming are increasingly referred to as plant-made pharmaceuticals (PMPs)

Plant derived pharmaceutical proteins

A typical Molecular Farming project

Plants that are used as host

Maíz, arroz, trigo, soja, tomate, patata, mostaza, nabo, alfalfa, banana, tabaco, algas verdes.

Which plant is the best?

• Tejidos verdes(Tabaco / Alfalfa)

Alfalfa and tobacco leaves can be cut several

times per year biomass 25-100 ton per ha (more protein)• Tobacco is easy to transform• Potatoes / Rice / Wheat• Seed is simple in component so protein can be purified

easily• Seed or tuber can be store for long time• Rice level of antibody did not decline significantly after

store at room temperature for 6 months• Potatoes lost antibody 50% after storage for 18 months

Porque plantas?

Las plantas tienen una habilidad natural para hacer proteínas humanas y animales.

Como se hace?

Cuales son las aplicaciones?

Molecular Farming 1 - Medical proteins

El primer acercamiento del molecular farming es la producción en masa de proteínas medicas de alto valor ( como por ejemplo: insulina, interferon, hormona del crecimiento, anticuerpos, proteínas de la sangre, etc.) aun bajo costo.

Molecular Farming 2 - Industrial enzymes

El segundo acercamiento al molecular farming es particularmente atractivo para la producción de proteínas cuando no es necesario un alto grado de purificación.

Molecular Farming 3 - Edible Vaccine

Las vacunas comestibles pueden ser usadas en lugar de la mayoría de las vacunas inyectadas.

Vacunas en plantas

Subunit vaccine

Specific macromolecules that induce a protective immune response against pathogen

Advantage: safer than traditional vaccine b/c only part is used

Disadvantage• Expensive to produce• Not heat stable

Advantage of plant-based vaccine production

• Simple: sunlight / mineral / water• Plant cell can correctly process proteins that are

complex• No contamination with animal pathogen• Heat stable environment (No need for

refrigerator)• Enable oral delivery (No pain and hazard of

injection)• Bio-encapsulation

History of antigen production in plants

PLANT DERIVED VACCINE• 1990 Streptococcus surface antigen in tobacco• Hepatitis B antigen in tobacco / lettuce• Rabies antigen in tomato• Cholera antigen in tobacco / potatoes• Human cytomegalovirus antigen in tobacco• Norwark virus antigen in potatoes• Foot and mouse disease in Arabidopsis

Clinical trial of edible vaccine in human

Transgenc Potatoes expressing part of toxin from E.coli

Treatment 50-100grTransgeni

c potatoes

Control normal

potatoes

10/11Immuneresponse

0/3No inmune

respose

VENTAJAS Y DESVENTAJAS

VENTAJAS• Las plantas no transportas patógenos que puedan se

perjudiciales para la salud humana.• No proteínas en plantas que sean similares a las

humanas.• Las plantas tienen relaciones cerradas con animales y

humanos los cuales pueden procesar y configurar correctamente sus proteínas.

DESVENTAJAS

• Molecular farming es una aplicación de la ingeniería genética.

Different steps to generate transgenic plants suitable for molecular farming of pharmaceuticals: (a) choice of a target gene and insertion into plant expression vector; (b) transfer of the expression construct into Agrobacterium tumefaciens; (c) plant transformation and selection under sterile condition; (d) generation of transgenic plants; (e) verification of protein accumulation, e.g. by Western blot, ELISA, microscopy; (f) functional proof in animal model, e.g. testing antibody formation after vaccination; (g) clinical studies; and (h) further development of the product and of the downstream processing technology as well as up scaling-up. P, promoter; G, gene; T, terminator.