Recombinant Plasmodium falciparum glutamate rich protein; purification and use in enzyme-linked immunosorbent assay.
A method for purification of a recombinant Plasmodium falciparum protein produced in E. coli and its use in an enzyme-linked immunosorbent assay (ELISA) is described. The cloned gene fragment encodes GLURP,489-1271 the carboxy-terminal 783 amino acid residue portion of a 1271 amino acid residue P. falciparum glutamate rich protein (GLURP), with a molecular weight of 220 kilodalton. The protein is associated with all parasite stages in the human host. Examination of sera from 105 adult Liberians living in a malaria endemic area revealed anti-GLURP IgG antibodies in 98% of the sera. The recombinant GLURP489-1271 was expressed as a chimeric protein, fused with E. coli beta-galactosidase. However, antibodies in sera were directed only against the malaria part of the fusion protein and not against beta-galactosidase. Antigen from in vitro P. falciparum cultures of isolates from Tanzania (F32), Papua New Guinea (MAD20) and Honduras (HB3) completely absorbed specific antibodies, indicating the presence of conserved epitopes produced by all isolates of P. falciparum. Recombinant GLURP489-1271 ELISA is sensitive and rapid, and therefore well-suited for sero-epidemiological studies, and for control of the immunogenicity of a possible future P. falciparum vaccine utilizing epitopes from GLURP.
Dziegiel
M
M
Statens Seruminstitut, Copenhagen, Denmark.
Borre
M B
MB
Jepsen
S
S
Hogh
B
B
Petersen
E
E
Vuust
J
J
D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D000918:Antibody Specificity; D002850:Chromatography, Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D010963:Plasmodium falciparum / Q000276:immunology*; D015800:Protozoan Proteins / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification
D000913:Antibodies, Protozoan; D015800:Protozoan Proteins; D011994:Recombinant Proteins