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Abstract
Cell Concentration(cells/ml)
6.00E+05
5.00E+05
4.00E+05
3.00E+05
2.00E+05
1.00E+05
0.00E+00
0
12 24 36 48 60 72 84 96 108 120
Time (hours)
Problem Statement
The current method used for production of ND vaccine is by
propagating the NDV in egg embryo, but this method is only sufficient
for small scale production of vaccine. To solve this problem, vaccine
production by propagating NDV virus in mammalian cell is the most
ideal method as it allowed easier upscaling for the process involved
with spinner flask or bioreactor.
Project Objectives
1. To propagate NDV in Vero cells for development of ND vaccines.
2. To determine the optimum condition needed for the virus
propagation in T-flask.
Research Methodology
Frozen Vero Cell line
was thawed and cultured
until passage 3 (p3)
NDV was infected on semi confluent p3 of
Vero cells until p5. On each passage, virus
culture was harvested, stored at 20oC and
infected again until it reached passage 5. (p5)
16 samples was taken
Block
Serum
Concentration (%)
MOI
(pfu/cells)
TCID50 (TCID50/ml)
Block 1
2.00
11.00
2.16 x 109
Block 1
2.00
11.00
1.48 x 109
Block 1
2.50
2.00
9.78 x 107
Block 1
1.50
11.00
2.58 x 109
Block 1
2.00
2.00
4.42 x 107
Block 1
2.50
20.00
3.97 x1010
Block 1
2.00
20.00
2.06 x 1010
Block 1
2.50
11.00
1.34 x109
Block 1
1.50
20.00
9.03 x 109
10
Block 1
2.00
11.00
3.07 x 109
11
Block 1
1.50
2.00
1.08 x107
Multiplicity
Virus
Infectivity
Concentration
of Infection
Titre (TCID50/ml)
2.48 %
20.00
2.547 x 1010
2.48 %
20.00
1.29 x 109
Selected
Solutions
Validation
Conclusion
1. All of the objectives stated in this project was successfully
achieved.
2. The difference in TCID50 response in the validation
experiment was due to possible human or technical error.
3. The optimum value gained for MOI was 20.00 and serum
concentration was 2.48%.
4. These result is an important information when large scale
production of NDV is considered.
Acknowledgment:
Special thanks to DR. Raha, Bro Azmir Ariffin and DR. Munirah for
the completion of this project.
Adapted from IRIE 2010 template