Expression of a Chlamydia anticarbohydrate single-chain antibody as a maltose binding fusion protein.
A single-chain antibody fragment has been constructed for an antibody that binds to the Chlamydia specific carbohydrate structure of the lipopolysaccharide. Single-chain protein was expressed and secreted into the periplasmic space of E. coli as a fusion protein with the maltose binding protein. The fusion protein was purified in one step by virtue of its ability to bind to maltose. In a sandwich ELISA, the eluted protein bound Chlamydia lipopolysaccharide, which demonstrates that the single-chain protein domain will function as part of a fusion protein. The expression of maltose binding fusion proteins into the periplasmic space could be used for production of other single-chain antibodies or protein fragments requiring appropriate folding and disulfide bond formation.
Malinowski
D P
DP
Department of Molecular Biology, Becton Dickinson Research Center, Triangle Park, NC 27709.
Gourley
M
M
Edelstein
S
S
Pearson
R E
RE
D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis* / Q000276:immunology* / Q000378:metabolism*; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000276:immunology*; D002689:Chlamydia / Q000276:immunology*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004220:Disulfides; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology / Q000378:metabolism; D008320:Maltose / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism*
D000907:Antibodies, Bacterial; D002241:Carbohydrates; D004269:DNA, Bacterial; D004220:Disulfides; D008070:Lipopolysaccharides; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D008320:Maltose