title: VAR2CSA domains expressed in Escherichia coli induce cross-reactive antibodies to native protein.
abstract: The variant surface antigen VAR2CSA is a pregnancy malaria vaccine candidate, but its size and polymorphism are obstacles to development. We expressed 3D7-type VAR2CSA domains in Escherichia coli as insoluble His-tagged proteins (Duffy binding-like [DBL] domains DBL1, DBL3, DBL4, and DBL5) that were denatured and refolded or as soluble glutathione S-transferase-tagged protein (DBL6). Anti-DBL5 antiserum cross-reacted with surface proteins of chondroitin sulfate A (CSA)-binding laboratory strains (3D7-CSA and FCR3-CSA) and a clinical pregnancy malaria isolate, whereas anti-DBL6 antiserum reacted only to 3D7 surface protein. This is the first report that E. coli-expressed VAR2CSA domains induce antibody to native VAR2CSA.
authors.affiliation: Seattle Biomedical Research Institute, University of Washington, Seattle, USA.
authors.forename: Andrew V
authors.initials: AV
authors.lastname: Oleinikov
authors.forename: Susan E
authors.initials: SE
authors.lastname: Francis
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Dorfman
authors.forename: Eddie
authors.initials: E
authors.lastname: Rossnagle
authors.forename: Stephanie
authors.initials: S
authors.lastname: Balcaitis
authors.forename: Tony
authors.initials: T
authors.lastname: Getz
authors.forename: Marion
authors.initials: M
authors.lastname: Avril
authors.forename: Severin
authors.initials: S
authors.lastname: Gose
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Smith
authors.forename: Michal
authors.initials: M
authors.lastname: Fried
authors.forename: Patrick E
authors.initials: PE
authors.lastname: Duffy
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000096:biosynthesis / Q000276:immunology; D000953:Antigens, Protozoan / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D004912:Erythrocytes / Q000469:parasitology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D016778:Malaria, Falciparum / Q000469:parasitology; D008969:Molecular Sequence Data; D010802:Phylogeny; D010963:Plasmodium falciparum / Q000235:genetics / Q000276:immunology*; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D015597:Pregnancy Complications, Parasitic / Q000469:parasitology; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D016415:Sequence Alignment
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D011994:Recombinant Proteins; C512601:VAR2CSA protein, Plasmodium falciparum