Scaph.net Presentation about viral marketing in healthcare, ICEA 2013 conference in Bucharest
Viral Nanoparticle Research Conference
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Transcript of Viral Nanoparticle Research Conference
![Page 1: Viral Nanoparticle Research Conference](https://reader036.fdocuments.net/reader036/viewer/2022062523/588114f31a28abb9388b47e1/html5/thumbnails/1.jpg)
MODIFIED BACTERIOPHAGE
LAMBDA PROCAPSIDS FOR USE AS A DRUG
DELIVERY PLATFORM.Presentation by:Nicole Rowley and Kyle Koshland
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◎ Advantages of using viruses
◎ What are Procapsids?
◎ Optimization of Procapsid Production
◎ Procapsid Modification
◎ Future Directions
Presentation Overview
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Advantages of Viruses◎ Variety of shapes and sizes in
nanometer size range
http://epidemiologiamolecular.com/wp-content/uploads/2011/08/clip_image0064.jpg
◎ Non-toxic◎ Inexpensive to produce◎ Amino acid residues can be
chemically and genetically modified
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◎ Procapsid without DNA or tail◎ Bacteriophage of E. coli
What Are Lambda Procapsids?
http://pdbj.org/emnavi/emnavi_detail.php?id=1507
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2 Main Goals:
1. Procapsid Prep
2. Procapsid Modification
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2 Main Goals:
1. Procapsid Prep
2. Procapsid ModificationFor drug
packaging
For quantification of modification
sites
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2 Main Goals:
1. Procapsid Prep
2. Procapsid Modification
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And the difficulties of finding a procedure that works!
1.Lambda Procapsid Prep
http://pdbj.org/emnavi/emnavi_detail.php?id=1507
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Plasmid TransformationDNA GpE/GpD Plasmid
Origin of Replication
Ampicillin Resistance
GpE/GpD
T7 Promotor
Operator
http://vignette2.wikia.nocookie.net/mmg-233-2013-genetics-genomics/images/e/e2/Transformation.png/revision/latest?cb=20131002205536
Heat Shock
LacI
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Procapsid Formation
http://viralzone.expasy.org/all_by_protein/3945.html
◎ Scaffolding proteins assist
○ Bonds break up completion
○ Not occurring in λ procapsid◎ Assemblies by Brownian Motion
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Ladd
er
+Con
trol
Resea
rch P
rep
SDS Gel Shows No ProductkDa
260
160
11080
60 50
40
3020
Capsid Protein
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Confirmation of Product
Ladd
er
+ Con
trol
Resea
rch P
rep
Capsid Protein
kDa260
160
110 80
60
50 40
30
20 1510
Ladder
Research Prep
kDa260
160
110
80
60
5040
30
20
Capsid Protein
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Intact Procapsid Confirmed
Intact
Denatured
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2 Main Goals:
1. Procapsid Prep
2. Procapsid Modification
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2 Main Goals:
1. Procapsid Prep
2. Procapsid Modification
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2.Procapsid Shell Modifications
https://www.carthage.edu/live/files/723-heidifenskemaxwell-maxhurickposterpdf
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Maleimide Modification
http://www.drugtargetreview.com/4442/content-type/industry-news/genticel-announces-positive-preclinical-proof-of-concept-results-for-its-multivalent-hpv-therapeutic-vaccine-gtl002/
R = Cy5
Intact
Denatured
Dye
Intact
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NHS Ester Modification
http://www.drugtargetreview.com/4442/content-type/industry-news/genticel-announces-positive-preclinical-proof-of-concept-results-for-its-multivalent-hpv-therapeutic-vaccine-gtl002/
R = Cy5
Intact Intact
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Quantification labels◎ Used Beer’s Law
○ Extinction coefficient◎ Average successful labeled amino acids
○ Lysines:
◉ 274 Cy5/Intact Procapsid
○ Cysteines:
◉ 208 Cy5/Intact Procapsid
http://www.pharmaceuticalonline.com/doc/lambert-beer-law-0001
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Quantification stability◎ Stability after conjugation
○ Lysine chemistry:
◉ 92.75% Intact○ Cysteine chemistry:
◉ 50.67% Intact◎ Calculated from area under curve
○ Ratio of intact to broken procapsids
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Further Experimentatio
n◎ Procapsid modifications
○ Attach linker and targeting protein
◎ Mouse studies
○ Inflammation in lupus mouse model
○ Blood-Brain barrier
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Any Questions?
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Thank You
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FPLC: Procapsid (Denatured)
FPLC Results