Methods and composition for secretion of heterologous polypeptides
AssigneeGenentech, Inc.
InventorsMatthew Marrichi, Dorothea E. Reilly
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Abstract
The present invention relates generally to the fields of molecular biology and protein technology. More specifically, the invention concerns signal sequences for the secretion of heterologous polypeptide from bacteria. The invention also concerns recombinant polypeptides and uses thereof.
Claims38
A method of making an a heterologous polypeptide, said method comprising culturing a host cell comprising polynucleotide comprising a variant translation initiation region (TIR) comprising nucleic acid variants of a PhoA, MalE, DsbA or STII secretion signal region operably linked to a polynucleotide encoding the heterologous polypeptide, so that the polynucleotide is expressed, whereby upon expression of the heterologous protein in a host cell, the heterologous polypeptide is folded to form a biologically active heterologous polypeptide.
A method of making an antibody, said method comprising culturing a host cell comprising a polynucleotide comprising (1) a first TIR operably linked to a polynucleotide encoding an antibody heavy chain, wherein the TIR comprises a co-translational prokaryotic secretion signal sequence;
- and (2) a second TIR operably linked to a polynucleotide encoding an antibody light chain, wherein the second TIR comprises a co-translational or post-translational prokaryotic secretion signal sequence, so that the antibody is expressed, whereby upon expression of the antibody in the host cell, the heavy and light chains are folded and assembled to form a biologically active antibody.
A method of secreting an antibody from a cell, said method comprising culturing a host cell comprising a polynucleotide comprising (1) a first TIR operably linked to a polynucleotide encoding an antibody heavy chain, wherein the TIR comprises a co-translational prokaryotic secretion signal sequence;
- and (2) a second TIR operably linked to a polynucleotide encoding an antibody light chain, wherein the second TIR comprises a co-translational or post-translational prokaryotic secretion signal sequence, so that the antibody is expressed and the antibody is secreted.
A method of translocating an antibody from a cell, said method comprising culturing a host cell comprising a polynucleotide comprises (1) a first TIR operably linked to a polynucleotide encoding an antibody heavy chain, wherein the TIR comprises a co-translational prokaryotic secretion signal sequence;
- and (2) a second TIR operably linked to a polynucleotide encoding an antibody light chain, wherein the second TIR comprises a co-translational or post-translational prokaryotic secretion signal sequence, so that the polynucleotide is expressed and the antibody is translocated
Dependencies are inferred from the claim text.