Purification, Characterization and Vaccine Potential of Trypanosoma brucei brucei Glycosyl Phosphatidyl Inositol Specific Phospholipase C.
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Date
2015
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Abstract
Aims: To investigate possible use of Glycosylphosphatidylinositol-specific phospholipase C (GPIPLC)
as a target protein for the development of vaccine against Trypanosoma brucei brucei
infection was investigated.
Study Design: GPI-PLC from T. brucei brucei was purified, characterized and the protein was
used as antigen in raising antibody against the parasite
Place and Duration: Department of Biochemistry, Ahmadu Bello University Zaria-Nigeria,
between September 2011 and October 2012
Methodology: GPI-PLC was isolated from T. brucei brucei and purified by ammonium sulphate
precipitation, gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE).
The GPI-PLC was further used to raise antisera in rabbits, which was subsequently used
to immunize rats for 14 and 21 days pre-infection to investigate the possible use of T. b. brucei GPI-PLC as target protein in vaccine production against T. b. brucei infection.
Results: An overall yield of 48.76% and purification fold of 10.86 were recorded after gel filtration.
The result from SDS-PAGE showed the enzyme to be a 39.585 kDa protein with optimum
temperature, optimum pH and activation energy to be 35°C, 8.1 and 19.494 kJ/ mol respectively.
The Vmax and Km values were 6.67 × 10-3 μmol/hr and 2.67 × 10-3 μM respectively when 212.5
μg of enzyme was used in the reaction mixture. Immunization with anti GPI-PLC for 14 and 21
days pre-infection significantly lowered the Packed Cell Volume (PCV). Result for the time course
of parasitemia following infection with 7.9 x 105 Cells/ml showed a decrease in parasitemia level,
thus leading to lowering of mortality rates in Groups immunized with GPI-PLC for 14 and 21 days
pre-infection by 20% and 40% respectively relative to Group infected but not treated.
Conclusion: These results suggest that GPI-PLC as a target protein significantly reduced the
progression of the T. b. brucei infection.
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Keywords
Purification, characterization, antigen, vaccine, T b, brucei, GPI-PLC
Citation
Abdullahi A S, Nzelibe H C, Atawodi S E. Purification, Characterization and Vaccine Potential of Trypanosoma brucei brucei Glycosyl Phosphatidyl Inositol Specific Phospholipase C. International Journal of TROPICAL DISEASE & Health. 2015; 5(1): 71-83.