This document provides an overview of biopharming, which is the use of plants to produce useful molecules for non-food applications. It discusses what biopharming is, why plants are used, current and evolving regulation, and risks and concerns. Specifically, it covers plant-made pharmaceuticals and industrial products, strategies for biopharming including plant expression systems and targeted tissues, examples of products on the market and in development, regulatory systems and guidelines, case studies, safeguard suggestions, alternatives, economics considerations, and directions for the future of this agricultural biotechnology.

1. AssignmentAssignment
Subject : Breeding Designer Crops (GPB822)Subject : Breeding Designer Crops (GPB822)
Presented by: Mr.Presented by: Mr. IndranilIndranil BhattacharjeeBhattacharjee
Student I.D. No.: 17PHGPB102Student I.D. No.: 17PHGPB102
Presented to : Prof. (Dr.)Presented to : Prof. (Dr.) B.G.SureshB.G.Suresh
SamSam HigginbottomHigginbottom University of Agriculture, Technology &University of Agriculture, Technology &
SciencesSciences
AllahabadAllahabad--211007211007

2. Lecture Outline
• What is biopharming?
• Why use plants?
• Current and evolving regulation
• What are the risks and concerns?

5. The use of agricultural plants for the production of
useful molecules for non food, feed or fiber
applications. (also called molecular farming,
pharming, or biopharming)
What is biopharming?

6. The use of agricultural plants for the production of
useful molecules for non food, feed or fiber
applications. (also called molecular farming,
pharming, or biopharming)
Plants are already grown to produce valuable
molecules, including many drugs.
Biopharming is different because the plants are
genetically engineered (GE) to produce the
molecules we want them to.
What is biopharming?

7. Plant Products
• Over 120 pharmaceutical products currently in
use are derived from plants. Mainly from tropical
forest species (e.g. Taxol from Yew trees)
1. Plant derived pharmaceuticals (non-GE)

8. Industrial products
• proteins
• enzymes
• modified starches
• fats
• oils
• waxes
• plastics
Pharmaceuticals
• recombinant human
proteins
• therapeutic proteins and
pharmaceutical
intermediates
• antibodies (plantibodies)
• vaccines
2. Plant-made pharmaceuticals (PMPs) and
industrial products (PMIP) (GE)
1.Plant-derived pharmaceuticals (non-GE)
Plant Products

9. Strategies for Biopharming
1.Plant gene expression strategies
• Transient transformation
• adv. – quick and easy production
• disadv. – small amount of product, processing pblms
• Stable transformation
• adv. – use for producing large quantities of protein, stability
and storage
• disadv – gene flow - outcrossing w/native species
• Chloroplast transformation
• adv. – reduce gene flow through pollen
• disadv. – protein not stable for long periods of time
therefore complications w/extraction/processing times

10. 1. Plant gene expression strategies
Protein quantity and preservation
• Whole plant
• adv. - an obtain large amts of protein
• disadv. - problems w/preservation
• examples - tobacco, alfalfa, duckweed
• Target specific tissues (e.g. seed, root)
• adv. - high amts of protein in seed/root, long-term
storage capability.
• examples: soy, corn, rice, barley
2. Location of transgene expression
Strategies for Biopharming

11. 1. Plant gene expression system
2. Location of trans-gene expression
3. Selection of plant species and characteristics
• Mode of reproduction – self/outcrossing
• Yield, harvest, production, processing
Strategies for Biopharming

13. Advantages
Cost reduction
- scalability (e.g. Enbrel® )
- low/no inputs
- low capital cost
Stability
- storage
Safety
- eukaroytic production system
- free of animal viruses (e.g.
BSE)
Why use plants?
Disadvantages
Environment
contamination
- gene flow
- wildlife exposure
Food supply
contamination
- mistaken/intentional mixing
w/human food
Health safety
concerns
- Variable, case-specific

14. Avidin by Sigma
• transgenic corn
• traditionally isolated from chicken egg whites
• used in medical diagnostics
GUS (b-glycuronidase) by Sigma
• transgenic corn
• traditionally isolated from bacterial
sources (E.Coli)
• used as visual marker in research labs
Trypsin by Sigma
• transgenic corn
• traditionally isolated from bovine pancreas
• variety of applications, including biopharmaceutical
processing
• first large scale transgenic plant product
• Worldwide market = US$120 million in 2004
Industrial products on the market

15. Industrial products close to market

16. • Plant- made vaccines (edible vaccines)
• Plant-made antibodies (plantibodies)
• Plant-made therapeutic proteins and
intermediates
Unlike PMIPs, no PMPS are currently available
on the market
Plant-made Pharmaceuticals (PMPs)

17. Edible vaccines
Advantages:
Administered directly
• no purification required
• no hazards assoc. w/injections
Production
• may be grown locally, where needed most
• no transportation costs
Naturally stored
• no need for refrigeration or special storage
Plant-made Vaccines

18. Examples of edible vaccines under development:
• pig vaccine in corn
• HIV-suppressing protein in spinach
• human vaccine for hepatitis B in potato
Plant-made Vaccines

19. • Plantibodies - monoclonal antibodies produced
in plants
• Plants used include tobacco, corn, potatoes,
soy, alfalfa, and rice
• Free from potential contamination of
mammalian viruses
• Examples: cancer, dental caries, herpes
simplex virus, respiratory syncytial virus
Plantibodies

20. Plantibodies
**GE Corn can produce up to 1 kg antibody/acre
and can be stored at RT for up to 5 years.
Humphreys DP et al. Curr Opin Drug Discover Dev 2001; 4:172-85.
Dental Caries MAb– expected to reach the market
soon
MAb directed against genital herpes – estimated to
reach market within 5 years
(Horn et al, 2004)

21. Therapeutic proteins and intermediates
• Blood substitutes – human hemoglobin
• Proteins to treat diseases such as CF, HIV,
Hypertension, Hepatitis B…..many others
Plant made Pharmaceuticals

22. **To date, there are no plant-produced
pharmaceuticals commercially available
Plant made Pharmaceuticals
Patient advocacy groups:
American Autoimmune Related Diseases Association
Arthritis Foundation
Cystic Fibrosis Foundation

23. Current ‘Pharm’ Companies

25. Current ‘Pharm’ Companies
•LEX System™
•Lemna (duckweed)
•trangenic tobacco
•PMPs and non-protein substances
(flavors and fragrances, medicinals,
and natural insecticides)
Kentucky Tobacco Research and
Development Center
Controlled Pharming Ventures
•collaboration w/Purdue
•transgenic corn
•converted limestone mine facility

26. Rhizosecretion
• Monoclonal antibodies
(Drake et al., 2003)
• Recombinant proteins
(Gaume et al, 2003)•biomass biorefinery
•based on switchgrass.
•used to produce PHAs
in green tissue plants
for fuel generation.
Current ‘Pharm’ Companies

27. • Genetically engineered Arabidopsis plants can
sequester arsenic from the soil. (Dhankher et al. 2002
Nature Biotechnology)
• Immunogenicity in human of an edible vaccine for
hepatitis B (Thanavala et al., 2005. PNAS)
Examples of Current Research
• Expression of single-chain antibodies in transgenic
plants. (Galeffi et al., 2005 Vaccine)
• Plant based HIV-1 vaccine candidate: Tat
protein produced in spinach. (Karasev et al. 2005
Vaccine)
• Plant-derived vaccines against diarrheal diseases.
(Tacket. 2005 Vaccine)

29. Environment contamination
• Gene flow via pollen
• Non-target species near field sites
e.g. butterflies, bees, etc
Food supply contamination
• Accident, intentional, gene flow
Health safety concerns
• Non-target organ responses
• Side-effects
• Allergenicity
Risks and Concerns

30. U.S. Regulatory System
(existing regulations)
Field Testing
-permits
-notifications
Determination of
non-regulated
status
Food safety
Feed safety
Pesticide and
herbicide
registration
USDA FDA EPA

31. Breakdown of Regulatory
System: Prodigene Incident 2002
2001 : Field trails of GE corn producing
pig vaccine were planted in IA and NE.
2002: USDA discovered “volunteer”
corn plants in fields in both IA and NE.
Soy was already planted in NE site.
$500,000 fine + $3 million to buy/destroy contaminated
soy

32. USDA Response to Incident
Revised regulations so that they were distinct from
commodity crops:
• Designated equipment must be used
• At least 5 inspections/yr
• Pharm crops must be grown at least 1 mile away
from any other fields and planted 28 days
before/after surrounding crops

33. FDA/USDA Guidance for Industry on Plant-Made
Pharmaceuticals Regulations
November 2004: Draft Document
Other challenges:
•Industrial hygiene and safety programs – these will
depend on the activity of the protein, route of exposure.
•Difficulty in obtaining relevant data because of high
species-specificity.
(Goldstein, 2005)
Current Evolving Regulations

34. www.ucsusa.org
Biopharming field trials in the US
Since 1995 ~ 300 biopharming plantings
The USDA receives/reviews applications for permits
for biopharm trials.

35. www.ucsusa.org
Biopharming field trials in the US
http://go.ucsusa.org/food_and_environment/pharm/index.php?s_keyword=XX
US Pharma Crop Database

36. Biopharming in Colorado

37. Biopharming in N.Carolina

38. The 2005 Oregon Biopharm Bill

39. Main concern is containment.
Opponents want:
• a guarantee of 0% contamination
of the food supply.
• full disclosure of field trials, crop,
gene, location, etc.
• an extensive regulatory framework
Biopharm opposition

40. 1. Physical differences
• e.g. “purple” maize, GFP
2. Sterility
• male sterile plants
• terminator technology
3. Easily detectable by addition of 'reporter
genes‘
• e.g. PCR markers
Suggested Safeguards for
biopharm operations

41. 4.Use chloroplast expression system
• will help increase yield
• will eliminate potential gene flow via pollen
• disadv. = technically difficult (Chlorogen
Company)
5. Complete disclosure of DNA sequences
6. Legislate for administration
Suggested Safeguards for
biopharm operations

42. Use only traditional drug production systems
•microbial, yeast and fungi
•mammalian cell culture
Use only fully contained production systems:
•plant cell cultures
•hydroponics (rhizosecretion)
•greenhouses
Use non-food crops
•tobacco
•hemp/cannabis
Alternatives to biopharming?

43. The expectation is for lower production costs
however there is no evidence that pharming
will produce cheaper, safe drugs.
Moreover, there are unknown costs associated
with containment, litigation and liability,
production…..others?
Economics

44. Future directions for
agricultural biotechnology?
Public perception
of risk
Regulation
Science has developed genetically enhanced crops and
has/can develop plant-made industrial and
pharmaceuticals crops.
The extent to which these crops will be further developed
for commercial and/or humanitarian use will ultimately
depend on…..