Cloning, expression and purification of Plasmodium knowlesi circumsporozoite protein and immunoblot analysis with P. knowlesi strain A1H1 protein extract
https://doi.org/10.47665/tb.39.2.004
- Author:
Tan, J.H.
1
;
Lau, Y.L.
1
Author Information
1. Department of Parasitology, Faculty of Medicine, Universiti Malaya, 50603 Kuala Lumpur, Malaysia
- Publication Type:Journal Article
- Keywords:
Circumsporozoite protein;
immunoblot analysis;
Plasmodium knowlesi;
recombinant protein expression.
- From:Tropical Biomedicine
2022;39(No.2):209-214
- CountryMalaysia
- Language:English
-
Abstract:
Circumsporozoite protein (CSP) is a sporozoite major surface protein of Plasmodium species. The protein
showed promising protection level as a vaccine candidate against Plasmodium falciparum infection.
There is a lack of studies on P. knowlesi CSP (PkCSP) as a vaccine candidate due to the high polymorphic
characteristic of central repeat region. Recent studies showed the protein has a relatively conserved
region at the C-terminal, which consists of T- and B-cell epitopes. This could be the target region for
vaccine development against the pre-erythrocytic stage of the parasite. In this study, recombinant
PkCSP was expressed using Escherichia coli system. Recombinant PkCSP was immunized in animal
models and the antiserum was evaluated using immunoblot analysis. Results showed that PkCSP can
be successfully expressed using the bacterial system. Endpoint titre of the antiserum were ranged up
to 1:819200. Immunoblot analysis showed the antiserum recognized recombinant PkCSP but not total
protein extract from P. knowlesi erythrocytic stage. In conclusion, PkCSP could elicit strong immune
response in animal models. However, serum antibodies could not recognize protein from the parasite’s
erythrocytic stage extract indicating it is not expressed at the erythrocytic stage. Therefore, PkCSP
remains as a potential pre-erythrocytic vaccine candidate against P. knowlesi infection.
- Full text:8.2022my1333.pdf
