what is viral nanoparticale application modification

1. 1

2. Nanoparticles
• Size: 1 to 100 nm
• Organic and inorganic in nature
• Metalic, liposomes and dendrimers etc.
• Used as carrier molecules
• Problems- toxic, non-degradable and non-specific
2

3. Viral nanoparticles - a versatile nanomachine
Saurav Saha
(2014-11-106)
Centre for Plant Biotechnology and Molecular Biology
College of Horticulture
3

4. Outline
• Introduction to virus
• Virus as a nanomachine
• Development of viral nanoparticles
• Applications
• Challenges
• Recent achievements
• Summary
• Conclusion
4

5. Introduction to virus
 Naturally occurring biomolecule
 Size- 15 to 2000 nm
 Rod-like or spherical in shape
 Capsid- outer protein coat
 Genomic material- DNA or RNA
 Deliver genome in host cells
5
(Grasso and Santi, 2010 )

6. • High strength of capsid protein
• Polyvalent and self-assembly process
• Monodisperse structure
• Mass production of viruses
6
(Alexander et al., 2013)
Virus as a nanomachine

7. 7
(Kristopher et al., 2015)
…virus as a nanomachine

8. Development of viral nanoparticle
8
Virus
Genetic
engineering
Bioconjugation Biomineralization Encapsulation
Modifications

9. • Epitope sequence
• Targeting sequence
• Unnatural amino acid
9
(Merzlyak et al., 2008)
Genetic engineering approach

10.  Coating of various substances within another material
 Assemble and de-assemble in-vitro
 Artificial polymer, enzyme, metallic nanoparticles
10
Encapsulation
(Carissa et al., 2010)

11.  pH > 6.5- swells and release RNA molecule
 pH decreases- PSS molecule assemble and entrapped
Polystyrene encapsulation
11
(Soto et al., 2010)

12. Biomineralization
Accumulation of minerals in cells and tissues
12
(Wang et al., 2012)

13. Contd…
13
BVSE- Biomineralized- based virus shell engineering
CAR- Coxsackie virus and adinovirus receptor
(Wang et al., 2012)

14. Bioconjugation
Two main strategies
 Standard bioconjugation chemistries
 Copper-catalyzed azide-alkyne cycloaddition
14
(Smith et al., 2013)

15. Standard bioconjugation chemistries
15(Smith et al., 2013)

16.  Azides and alkynes in the presence of copper
 Popularly known as click reaction
 Conjugation of protein building block
Copper-catalyzed azide-alkyne cycloaddition
16(Smith et al., 2013)

17. Application of viral nanoparticles
17
Viral nanoparticles
(VNP)
Targeted drug
delivery
Vaccines Imaging
Plant disease
management

18. Targeted drug delivery
 Reduce drug toxicity and degradation
 Target particular organ
 Increase bioavailability and circulation
18
( Nicholas et al., 2014)

19.  Anti-cancer drug
 Poor selectivity
 High cardio toxicity
Doxorubicin (DOX) delivery
19
(Zeng et al., 2013)
FA- Folic acid

20. Cytotoxicity of various DOX formulations at
different DOX concentration
20( Zeng et al., 2013)

21. 0
10
20
30
40
50
60
70
80
90
Dox CMV-DOX CMV-FA-DOX
Apoptoticpercentage
 DOX concentration (5 µg/ml) incubate for 3 hr
Effect of DOX on cardiomyocytes cells
21
( Zeng et al., 2013 )

22. Vaccine Production
Virus like particles (VLPs)
Antigen stability
Potential to carry two or more different antigen (chimera)
22( Kristopher et al., 2015)

23. Flock house virus VLP production
23
( Destito et al., 2009 )

24. Universal influenza vaccine from VLP
 Globally 250,000–500,000 deaths annually
 Virus continually evolving
 H1, H2, H5, H6, H7, H10, and H11 hemagglutinin subtypes
 Universal vaccine are most effective
24
(Kang et al., 2009)

25. .…universal influenza vaccine from VLP
 Mice vaccinated with mixture of 1.5g each of H1, H3, H5, and
H7 VLPs
 Mice were boosted at 21 days post immunization
 After 35 days of immunization
 Mice are infected with different strains of virus
25
(Kang et al., 2009)

26. Effect of homologous challenge
26(Kang et al., 2009)
Bodyweight(%)
Survival(%)(Days)
H1N1

27. Effect of heterosubtypic challenge
27
Bodyweight(%)
Survival(%) Days
H6N1
(Kang et al., 2009)
Days

28. VLP type Antigen Indication Product
name
Status Reference
Non-
enveloped
Virus structural
protein
HBV GenHevac B® Licensed Soulie et al.,
1991
Non-
enveloped
Virus structural
protein
HPV Cervarix® Licensed Agnandji et
al., 2012
Non-
enveloped
(chimeric)
Parasite protein Malaria RTS,S Phase 1 El-Attar et
al.,
2009
Enveloped
(virosome )
Parasite protein Malaria PEV3 Phase 1/2 Cech et al.,
2011
HBV-Hepatitis B virus
HPV-Human papilloma virus
Other types of VLP based vaccine
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29. Vaccine for tumor/ cancer
29
 Tn glycan over expressed
 Low immunogenicity of carbohydrates (Tn)
 Induce a strong T cell-dependent immune
 Cowpea mosaic virus (CMV) conjugate with Tn glycoprotein
( Miermont et al., 2008 )

30. 0
2000
4000
6000
8000
10000
12000
14000
16000
day 0 day35 day 35+
GAlNac
day 35
comtrol
Titre
ELISA titres of anti-Tn conjugate with CMV
30
(GAINac- Tn antigen) ( Miermont et al., 2008 )

31. control IgG control IgM
Series1 10000 5000
0
2000
4000
6000
8000
10000
12000Titre
Titre of different types antibodies
31( Miermont et al., 2008 )

32.  Visual representation of internal structures
 Synthetic dye ,quantum dot and green fluorescent protein
 Low sensitive and photo stability
 VNP used as carrier for imaging dye
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Imaging
( Yoo et al., 2012)

33. Intravital vascular imaging
Fd- Florescine dextran
A4d- A488 dextran
Rhl- Rhadomine-leveled
33
(Lewis et al., 2006)

34. Effect of injection of viral nanoparticle based
fluorescence dye on mouse
34(Lewis et al., 2006)

35. In -vivo stability of fluorescent viral
nanoparticles
35
( Yoo et al., 2012)

36.  Worldwide crop damage - 157 million dollar
 Highly toxic contact and fumigant nematicides
 Abamectin (Abm)- biologically active compound
 Immobile in soil and photo-oxidative in nature
 VNPs used as carrier for abamectin
Plant parasitic nematode control
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37. Red clover necrotic mosaic virus (RCNMV) as VNP
37( Richard et al., 2015)

38. Performance of VNP loaded abamectin
Time (Hour)
pH 5.2
A- Within 5 hours 95% of the chemical diffuse out
B- Within 5 hours 25% of the chemical diffuse out
38( Richard et al., 2015)
pH 5.2
A B

39. Effect of VNP loaded abamectin on tomato
seedling
39Gall No gall ( Richard et al., 2015)

40. Recent achievements
40

41. • Herpes virus- genetically modified
• BioVex company in USA
• FDA approval for treating cancer
• Brand name imlygic
Cancer-hunting virus
41
(Bell et al., 2015)

42. Carbon-free hydrogen fuel
 Hydrogenase enzyme
 Protons (H+) and electrons (e-) into molecules of H2
 P22 bacteriophage coat protein to encapsulate
 Mixed with Protons and electrons-ferrying molecules
 Hydrogen produced inside a virus
42
( Robert et al., 2015 )

43. Challenges
• Purity in compound
• Encapsulate contaminants
• Manufacturing- not scalable or cost-effective
• Structural complexity
• Baculovirus as a vector
43
(Crisci et al., 2012 )

44. Summary
 Viral nanoparticles
 Features of viral nanoparticles
 Modification strategies
 Targeted drug delivery, vaccination and imaging
 Plant disease control
 Major Challenges
44

45. • VNPs used as biological nanocarrier
• Enormous application in biomedical and agriculture
• Modification- chemical and genetic
• Drug, toxin and targeted sequence
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Conclusion

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“Where Nature finishes producing its own species, man
begins, using natural things and with the help of this
nature, to create an infinity of species”
Leonardo-Da-Vinci

47. 1/17/2016 47
Thank you