The Generation of Adenovirus Vectors by ATPS Intended for Gene Therapy of CancerSAQLAIN SULEMAN

Introduction• Background information

Experiments

• Method• Results

Discussion

•Conclusions•Future work

Adenovirus life cycle

Genome

ATPS

Method

Growing adenovirus

Purifying adenovirus

Quantification

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

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

Results

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

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

Conclusions

Unsuccessful quantification of adenovirus vectors

PEG600- ammonium sulphate

ATPS:- Time effective- Cost effective- High yields

Acknowledgments

Dr M. Themis Dr P. Hewitson & Dr S. Ignatova Hassan Khonsari Dr E. Makarov Ataun Bhatti