viral nanoparticles
TRANSCRIPT
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WELCOME
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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
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Viral nanoparticles - a versatile nanomachine
Saurav Saha(2014-11-106)
Centre for Plant Biotechnology and Molecular BiologyCollege of Horticulture
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Outline
• Introduction to virus
• Virus as a nanomachine
• Development of viral nanoparticles
• Applications
• Challenges
• Recent achievements
• Summary
• Conclusion 4
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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
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(Grasso and Santi, 2010 )
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• High strength of capsid protein
• Polyvalent and self-assembly process
• Monodisperse structure
• Mass production of viruses
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(Alexander et al., 2013)
Virus as a nanomachine
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(Kristopher et al., 2015)
…virus as a nanomachine
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Development of viral nanoparticle
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Virus
Genetic engineering Bioconjugation Biomineralization Encapsulation
modifications
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• Epitope sequence
• Targeting sequence
• Unnatural amino acid
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(Merzlyak et al., 2008)
Genetic engineering approach
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Coating of various substances within another material
Assemble and de-assemble in-vitro
Artificial polymer, enzyme, metallic nanoparticles
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Encapsulation
(Carissa et al., 2010)
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pH > 6.5- swells and release RNA molecule
pH decreases- PSS molecule assemble and entrapped
Polystyrene encapsulation
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(Soto et al., 2010)
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Biomineralization
Accumulation of minerals in cells and tissues
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(Wang et al., 2012)
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Contd…
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BVSE- Biomineralized- based virus shell engineering
CAR- Coxsackie virus and adinovirus receptor (Wang et al., 2012)
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Bioconjugation
Two main strategies
Standard bioconjugation chemistries
Copper-catalyzed azide-alkyne cycloaddition
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(Smith et al., 2013)
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Standard bioconjugation chemistries
15(Smith et al., 2013)
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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)
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Application of viral nanoparticles
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Viral nanoparticles (VNP)
Targeted drug delivery Vaccines Imaging Plant diseases
management
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Targeted drug delivery
Reduce drug toxicity and degradation
Target particular organ
Increase bioavailability and circulation
18( Nicholas et al., 2014)
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Anti-cancer drug
Poor selectivity
High cardio toxicity
Doxorubicin (DOX) delivery
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(Zeng et al., 2013)FA- Folic acid
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Cytotoxicity of various DOX formulations at different DOX concentration
20( Zeng et al., 2013)
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Dox CMV-DOX CMV-FA-DOX0
102030405060708090
Apop
totic
per
cent
age
DOX concentration (5 µg/ml) incubate for 3 hr
Effect of DOX on cardiomyocytes cells
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( Zeng et al., 2013 )
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Vaccine Production
Virus like particles (VLPs)
Antigen stability
Potential to carry two or more different antigen (chimera)
22( Kristopher et al., 2015)
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Flock house virus VLP production
23( Destito et al., 2009 )
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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
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(Kang et al., 2009)
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.…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
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(Kang et al., 2009)
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Effect of homologous challenge
26(Kang et al., 2009)
Body
wei
ght (
%)
Surv
ival
(%)
(Days)
H1N1
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Effect of heterosubtypic challenge
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Body
wei
ght (
%)
Surv
ival
(%)
Days
H6N1
(Kang et al., 2009)
Days
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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 virusHPV-Human papilloma virus
Other types of VLP based vaccine
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Vaccine for tumor/ cancer
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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 )
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day 0 day35 day 35+ GAlNac
day 35 comtrol
0
2000
4000
6000
8000
10000
12000
14000
16000Ti
tre
ELISA titres of anti-Tn conjugate with CMV
30(GAINac- Tn antigen) ( Miermont et al., 2008 )
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control IgG control IgM0
2000
4000
6000
8000
10000
12000Ti
tre
Titre of different types antibodies
31( Miermont et al., 2008 )
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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)
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Intravital vascular imaging
Fd- Florescine dextranA4d- A488 dextranRhl- Rhadomine-leveled
33(Lewis et al., 2006)
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Effect of injection of viral nanoparticle based fluorescence dye on mouse
34(Lewis et al., 2006)
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In -vivo stability of fluorescent viral nanoparticles
35( Yoo et al., 2012)
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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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Red clover necrotic mosaic virus (RCNMV) as VNP
37( Richard et al., 2015)
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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
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Effect of VNP loaded abamectin on tomato seedling
39Gall No gall ( Richard et al., 2015)
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Recent achievements
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• Herpes virus- genetically modified
• BioVex company in USA
• FDA approval for treating cancer
• Brand name imlygic
Cancer-hunting virus
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(Bell et al., 2015)
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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
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( Robert et al., 2015 )
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Challenges
• Purity in compound
• Encapsulate contaminants
• Manufacturing- not scalable or cost-effective
• Structural complexity
• Baculovirus as a vector
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(Crisci et al., 2012 )
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Summary
Viral nanoparticles
Features of viral nanoparticles
Modification strategies
Targeted drug delivery, vaccination and imaging
Plant disease control
Major Challenges
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05/01/2023
• 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
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Thank youThank youThank you