D. Schnappinger et al., EXTRACELLULAR EXPRESSION OF NATIVE HUMAN ANTILYSOZYME FRAGMENTS IN STAPHYLOCOCCUS-CARNOSUS, FEMS microbiology letters, 129(2-3), 1995, pp. 121-127
Xylose-inducible vectors have been constructed for extracellular produ
ction of antibody fragments in Staphylococcus carnosus. The pre-pro se
quence of S. hyicus lipase was taken as secretional signal sequence, a
nd 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 rem
oval 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 ve
ctors. The pro sequence could be removed from the expression product b
y IgA protease treatment. When the light chain and the Fd fragment wer
e co-secreted as a protein fusion they accumulated in a structure capa
ble of heterodimerization after IgA cleavage. This fusion contains the
pre-pro sequence followed by the light chain, a second IgA site and t
he Fd fragment.