[Expression, isolation and purification of antibody fragments fused to maltose-binding protein in Escherichia coli].
We have fused the variable domains of a mouse antibody to the C-terminal end of the maltose-binding protein (malE), at the genetic level. The hybrid proteins were expressed in E. coli under control of the malEp promoter, and exported to the periplasm, at low temperature. They were purified by affinity chromatography on cross-linked amylose. When the two variable domains were fused together through a peptide link, the hybrid displayed similar affinity and specificity to the antigen as the native antibody.
Brégégère
F
F
Unité de Biochimie cellulaire, C.N.R.S.-U.A. n. 1129, Institut Pasteur, Paris.
Bedouelle
H
H
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D006824:Hybridization, Genetic / Q000235:genetics; D007128:Immunoglobulin Fragments / Q000235:genetics* / Q000302:isolation & purification; D008320:Maltose / Q000378:metabolism*; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics*
D002352:Carrier Proteins; D007128:Immunoglobulin Fragments; D011993:Recombinant Fusion Proteins; D008320:Maltose
Expression, exportation et purification de fragments d'anticorps fusionnés à la protéine affine du maltose d'Escherichia coli.