Eukaryotic signal sequences for polypeptide expression and polypeptide display libraries
View Patent ↗The present invention generally relates to methods and compositions for expressing proteins or polypeptides in prokaryotic hosts using eukaryotic signal sequences.
1. A method of displaying a polypeptide from a replicable genetic package, which method comprises the steps of expressing in a bacterial host cell a polypeptide display vector comprising a polynucleotide encoding the polypeptide, wherein the polynucleotide is operably linked to a eukaryotic signal sequence which encodes a signal peptide MRTLAILAAILLVALQAQA (SEQ ID NO: 2), whereby the polynucleotide sequence and signal sequence are expressed and secreted, and the expressed signal peptide sequence is processed from the expressed polypeptide, such that the polypeptide is displayed from the outer surface of the replicable genetic package, wherein the replicable genetic package is a bacteriophage or phagemid, and wherein the polypeptide is an antibody.
2. A method according to claim 1 , wherein the antibody is a mouse antibody, a human antibody, or a chimeric antibody.
3. A method according to claim 1 , wherein the antibody is a whole antibody or binding fragment thereof.
4. A method according to claim 1 , wherein the antibody is an antibody binding fragment that is a single chain fragment, an Fv fragment or a Fab fragment.
5. A method according to claim 1 , wherein the signal sequence is 5′ to the polynucleotide sequence encoding the antibody.
6. A method according to claim 1 , wherein the signal sequence and the antibody are separated by zero amino acids.
7. A method according to claim 1 , wherein the vector is a polycistronic vector.
8. A method according to claim 1 , wherein the bacteriophage is a filamentous phage.
9. A method according to claim 8 , wherein the filamentous phage is an M13, Fd or F1 phage.
10. A method according to claim 1 , further comprising the step of screening for binding of the antibody to a target of interest.