final year project presentation.2
TRANSCRIPT
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The Generation of Adenovirus Vectors by ATPS Intended for Gene Therapy of CancerSAQLAIN SULEMAN
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Introduction• Background information
Experiments
• Method• Results
Discussion
•Conclusions•Future work
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Adenovirus life cycle
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Genome
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ATPS
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Method
Growing adenovirus
Purifying adenovirus
Quantification
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HEK293T cell density
Estimated cell density of subsequent plates
2.0 x 106 1.0 x 106
9.875 x 106
2.46875 x 106
ResultsHeLa cell density
Estimated cell density on subsequent plates
8.125 x 106
2.03125 x 106
6.975 x 106
2.325 x 106
4.2625 x 106
1.065625 x 106
HepG2 cell density
Estimated cell density on subsequent plates
7.0 x 104
1.75 x 104
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Results
ATPS 1 ATPS 2 ATPS 3
Phase system Phase ratio
Phase one: PEG1000 – potassium phosphate
1: 1.17
Phase two: PEG8000 – potassium phosphate
1: 1.38
Phase three: PEG400 – ammonium sulphate
1: 2.57
Phase four: UCON970 – sodium citrate
1: 3.25
Phase five: PEG600 – ammonium sulphate
1: 1.67
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Results
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Results
Phase 1 Phase 2 Phase 3 Phase 40
10
20
30
40
50
60
70
80
90
100
UpperLower
Perc
enta
ge o
f Inf
ecte
d C
ells
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Results
Phase 1 Phase 2 Phase 30
10
20
30
40
50
60
70
Upper PhaseLower Phase
Perc
enta
ge o
f in
fect
ed c
ells
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Conclusions
Unsuccessful quantification of adenovirus vectors
PEG600- ammonium sulphate
ATPS:- Time effective- Cost effective- High yields
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Acknowledgments
Dr M. Themis Dr P. Hewitson & Dr S. Ignatova Hassan Khonsari Dr E. Makarov Ataun Bhatti