Presentation on Plantibodies and their types.pdf

It contains detailed about Plantibodies and their types, advantages and disadvantage and their application

1. VIVEKANANDHA ARTS AND SCIENCE COLLEGE
FOR WOMEN , SANKAGIRI
DEPARTMENT OF MICROBIOLOGY
SUBJECT : SOIL AND ENVIRONMENTAL MICROBIOLOGY
TOPIC : PLANTIBODIES
SUBJECT INCHARGE :
Dr.R.DINESHKUMAR ,
ASSISTANT PROFESSOR ,
DEPARTMENT OF MICROBIOLOGY ,
VIAAS – SANKAGIRI.
SUBMITTED BY :
V. ABINAYA ,
II – MSC MICROBIOLOGY ,
VIAAS – SANKAGIRI .

2. CONTENTS
•Introduction
•Definition
•History
•Antibody Produced From Plants
•Components of a Plantibody Gene
•Types of Plantibody
•Methods for Plantibody Production
•Purification of Plantibody
•Techniques for Plantibody Production
•Advantage
•Applications
•Conclusion

3. INTRODUCTION
•The process involves introducing the entire Ig
gene encoding the desired antibody into the plant's
genome using techniques used to create transgenic
organisms.
•Plantibodies are antibodies produced in genetically modified plants
that express animal antibody genes.

4. DEFINITION OF PLANTIBODY
•Plantibodies are antibodies produced by plants
that have been genetically engineered with
animal DNA that encodes a specific human
antibody.

5. HISTORY OF PLANTIBODY
•The term "Plantibody" was coined around 1990 to describe antibodies
produced by plants that have been genetically engineered with animal
DNA.
•In 2000s - Plantibody production expands to other plant species (e.g.,
corn, soybeans, alfalfa)
•First Plantibody-based drug, Taliglucerase alfa (Elelyso), is approved
for Gaucher's disease (2006).

6. ANTIBODIES PRODUCED FROM PLANTS
•Guy's 13 : An anti-viral antibody produced in tobacco plants.
•2G12 - An anti-HIV antibody produced in tobacco plants.
• M18 : An anti-rabies antibody produced in tobacco plants.
•Taliglucerase alfa : A plant-made enzyme replacement therapy
for Gaucher's disease. Elelyso : A plant-made enzyme
replacement therapy for Gaucher's disease.
•Plant are being used as antibody factories, using their
endomembrane and secretory system to produce large amount of
clinically visible proteins.

7. COMPONENTS OF PLANTIBODY GENE
•Plantibodies gene construct can contain sequence for fragment (Fab,
scFv, whole length antibody).
•Effective promoters for increased expression of proteins attached
upstream of the genes. Eg. Hsp70 promoter is more effective the 35S
promoter.
•Signal peptide sequence - needed for targeting of antibody into ER for
proper folding of protein.
•C-terminal peptide- usually KDEL added to C-terminal of the peptide
that can increase expression and retention of the protein in cell.

8. TYPES OF PLANTIBODIES
Expression of
full-length
antibody
Expression of
antibody
fragments (Fab)
Expression of single
chain scFv or single
domain (VL) gones

9. TYPES OF PLANTS USED FOR PLANTIBODY
PRODUCTION
•Tobacco
•Corn
•Soybeans
•Alfalfa
•Arabidopsis
•CaroRIT : A plant made antibody againts ricin toxin.
•Zmapp : An Ebola treatment made from antibodies produced in
Tobacco Plants.

10. METHODS FOR PLANTIBODY
PRODUCTION
•Using transformation and transient expression to introduce new genes
into a host cell.
•Targeting to the APOPLASM by the tagging with a small peptide
sequence.
•Signal molecules added to target the protein to the Endoplasmic
Reticulum (E.R); to ensures correct folding of the protein.
•The Plantibodies are then modified by intrinsic plant mechanisms
(N-glycosylation).

11. •Higher protein levels obtained in the E.R and the apoplasm as
compared to the cytosol.

12. GLYCOSYLATION
•Glycosylation refers to the enzymatic process
that attaches glycans to proteins, lipids, or other
organic molecules.
• It occurs in all higher eukaryotes in the Golgi
complex.
•Glycans serve a variety of structural and
functional roles in membrane and secreted
proteins.
Fig ; N - glycan processing in plants

13. •Glycosylation by plants differs to that found in mammalian cells.
•Plant glycans are smaller and have different terminal sugar residues.
•It is thought this may have an immunogenic effect and have low
antigen-binding affinity
Glycosylation in Golgi
Human Plant

14. PURIFICATION OF PLANTIBODIES
•Plantibodies can be purified cheaply in large quantities.
•Transgenic seeds assure excellent storage properties and due to limited
range of endogenous proteins in seeds, separation of plantibody is less
complicated.
•There is no risk of spreading animal diseases to humans as the antibodies
are produced by plants.
•The production of large amounts of clinically viable protein especially
IgA has many applications in medicine.

15. 1.Extraction : Plant tissues are extracted to release the plantibody into a
solution.
2. Clarification : The extract is clarified to remove plant debris and
contaminants.
3.Protein precipitation : Proteins are precipitated using salts, solvents, or
polymers to concentrate the plantibody.
4.Chromatography : Affinity chromatography : Uses specific ligands to
bind and separate the plantibody.
5.Filtration : Removes impurities and concentrates the plantibody.

16. 6. Dialysis: Removes small molecules and buffer exchange.
7. Concentration :Concentrates the purified plantibody.
8. Formulation : Prepares the purified plantibody for storage, shipping, or
application.

17. TECHNIQUES FOR PLANTIBODY PRODUCTION
1. Agrobacterium-mediated transformation
2. Biolistics
3. Electroporation
4. Microinjection
5.CRISPR/Cas9 gene editing
6. RT-PCR and qRT-PCR
7. Western blotting and ELISA
8.HPLC and FPLC
9. Mass spectrometry
10. N-glycan analysis.
•Some specific techniques used in plantibody technology include ;

18. ADVANTAGES OF PLANTIBODIES
•Low production cost
•High scalability
•Easy purification
• Long shelf life
•Low risk of human pathogens
•Flexibility
•High yield
•Glycoengineering.
•Thermostability

19. APPLICATIONS OF PLANTIBODIES
•Plantibodies have various application such as ,
•Therapeutics
•Diagnosticts
•Vaccines
•Research tools
•Gene Expression Analysis
•Cosmetics

20. CONCLUSION
Genetically engineered plants acts as most desirable and
promising host for expression of Plantibodies for humans.
They stably express antibodies and its fragments and edible
vaccines in their edible tissues which are of therapeutic
importance.
High scalability, low-cost and safety characteristics present an
attractive approach for commercialization of pharmaceutical
products for the developing world.

21. THANK YOU