title: Extracellular expression of native human anti-lysozyme fragments in Staphylococcus carnosus.
abstract: Xylose-inducible vectors have been constructed for extracellular production of antibody fragments in Staphylococcus carnosus. The pre-pro sequence of S. hyicus lipase was taken as secretional signal sequence, and the S. xylosus Xyl repressor was used to confer xylose inducibility of transcription. Cleavage sites for the IgA protease were engineered between the pre-pro sequence and the antibody fragments to permit removal of the pro sequence. Extracellular expression of the light chain and the Fd fragment of a chimeric Fab fragment containing the variable regions of the anti-lysozyme antibody D1.3 was achieved with these vectors. The pro sequence could be removed from the expression product by IgA protease treatment. When the light chain and the Fd fragment were co-secreted as a protein fusion they accumulated in a structure capable of heterodimerization after IgA cleavage. This fusion contains the pre-pro sequence followed by the light chain, a second IgA site and the Fd fragment.
authors.affiliation: Institut für Mikrobiologie, Biochemie und Genetik der Friedrich-Alexander Universität Erlangen-Nürnberg, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Schnappinger
authors.forename: W
authors.initials: W
authors.lastname: Geissdörfer
authors.forename: C
authors.initials: C
authors.lastname: Sizemore
authors.forename: W
authors.initials: W
authors.lastname: Hillen
mesh_terms: D000906:Antibodies / Q000235:genetics; D001483:Base Sequence; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006801:Humans; D008969:Molecular Sequence Data; D009113:Muramidase / Q000276:immunology; D010957:Plasmids / Q000235:genetics*; D013210:Staphylococcus / Q000235:genetics*
chemical_list: D000906:Antibodies; D009113:Muramidase