Expression of an immunologically reactive merozoite surface protein (MSP-1(42)) in E. coli.
The 42-kDa carboxyl-terminal processing fragment of Plasmodium falciparum merozoite surface protein-1 (PfMSP-1(42)) is one of the anti-malarial vaccine candidate antigens. In the present study, recombinant MSP-1(42) was expressed as a fusion protein in a novel E. coli host. The average yield of the recombinant protein was 48 mg/l of bacterial culture. The antigenicity and immunogenicity of the purified protein were evaluated by comparing the results with those obtained from a well-characterized recombinant MSP-1(42) (Bmp42) expressed in the baculovirus expression system previously described from our laboratory. We observed that there is a high degree of similarities between the two recombinant proteins. Based on the results from T and B cell response, in vitro parasite growth inhibition, as well as cross-reactivities with several well-characterized MSP-1 specific Mabs, the bacterial expressed protein is apparently comparable to Bmp42 in terms of immunoreactivities. Our results suggest that the bacterial expression system could be employed to express immunologically active recombinant MSP-1(42) at elevated levels. This system may be an attractive alternative for producing a protective vaccine for human use at lower cost.
Leung
W H
WH
Department of Biochemistry, Chinese University of Hong Kong, Shatin, Hong Kong.
Meng
Z Q
ZQ
Hui
G
G
Ho
W K K
WK
D000818:Animals; D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan / Q000276:immunology; D000953:Antigens, Protozoan / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics; D049109:Cell Proliferation; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000469:parasitology; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D008213:Lymphocyte Activation; D017780:Malaria Vaccines / Q000276:immunology; D020066:Merozoite Surface Protein 1 / Q000235:genetics* / Q000276:immunology*; D051379:Mice; D010446:Peptide Fragments / Q000276:immunology; D010963:Plasmodium falciparum / Q000235:genetics*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D017780:Malaria Vaccines; D020066:Merozoite Surface Protein 1; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic