Recombinant Plasmodium falciparum glutamate rich protein; purification and use in enzyme-linked immunosorbent assay
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Recombinant Plasmodium falciparum glutamate rich protein; purification and use in enzyme-linked immunosorbent assay. / Dziegiel, Morten Hanefeld; Borre, M B; Jepsen, S; Hogh, B; Petersen, E; Vuust, J.
In: American Journal of Tropical Medicine and Hygiene, Vol. 44, No. 3, 03.1991, p. 306-13.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - Recombinant Plasmodium falciparum glutamate rich protein; purification and use in enzyme-linked immunosorbent assay
AU - Dziegiel, Morten Hanefeld
AU - Borre, M B
AU - Jepsen, S
AU - Hogh, B
AU - Petersen, E
AU - Vuust, J
PY - 1991/3
Y1 - 1991/3
N2 - 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.
AB - 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.
KW - Animals
KW - Antibodies, Protozoan
KW - Antibody Specificity
KW - Chromatography, Gel
KW - Enzyme-Linked Immunosorbent Assay
KW - Humans
KW - Plasmodium falciparum
KW - Protozoan Proteins
KW - Recombinant Proteins
M3 - Journal article
C2 - 2035752
VL - 44
SP - 306
EP - 313
JO - Journal. National Malaria Society
JF - Journal. National Malaria Society
SN - 0002-9637
IS - 3
ER -
ID: 47556425