Extracellular expression of native human anti-lysozyme fragments in Staphylococcus carnosus.

TitleExtracellular expression of native human anti-lysozyme fragments in Staphylococcus carnosus.
Publication TypeJournal Article
Year of Publication1995
AuthorsSchnappinger D, Geissdörfer W, Sizemore C, Hillen W
JournalFEMS Microbiol Lett
Date Published1995 Jun 15
KeywordsAntibodies, Base Sequence, Gene Expression, Gene Transfer Techniques, Humans, Molecular Sequence Data, Muramidase, Plasmids, Staphylococcus

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.

Alternate JournalFEMS Microbiol Lett
PubMed ID7607392

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