Polypeptide display libraries and methods of making and using thereof
First Claim
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1. A composition comprising:
- a fusion protein, the fusion protein comprisinga bacterial outer membrane protein,wherein the bacterial outer membrane protein is OmpA or OmpX,wherein a native N-terminus and a native C-terminus of the bacterial outer membrane protein are joined by a linker, andwherein the bacterial outer membrane protein comprises a non-native N-terminus and a non-native C-terminus derived from an extracellular loop of the bacterial outer membrane protein; and
a passenger polypeptide linked or fused directly to the non-native N-terminus or the non-native C-terminus.
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Abstract
Disclosed herein are expression vectors which display a passenger polypeptide on the outer surface of a biological entity. As disclosed herein the displayed passenger polypeptide is capable of interacting or binding with a given ligand. Also disclosed are methods of making and using the expression vectors. N/C terminal fusion expression vectors and methods of making and using are also disclosed.
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32 Claims
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1. A composition comprising:
a fusion protein, the fusion protein comprising a bacterial outer membrane protein, wherein the bacterial outer membrane protein is OmpA or OmpX, wherein a native N-terminus and a native C-terminus of the bacterial outer membrane protein are joined by a linker, and wherein the bacterial outer membrane protein comprises a non-native N-terminus and a non-native C-terminus derived from an extracellular loop of the bacterial outer membrane protein; and a passenger polypeptide linked or fused directly to the non-native N-terminus or the non-native C-terminus. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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18. A polypeptide display library comprising:
a plurality of host cells, each member of the plurality comprising a fusion protein disposed in an outer membrane of a host cell, the fusion protein comprising a bacterial outer membrane protein, wherein the bacterial outer membrane protein is OmpA or OmpX, wherein a native N-terminus and a native C-terminus of the bacterial outer membrane protein are joined by a linker, and wherein the bacterial outer membrane protein comprises a non-native N-terminus and a non-native C-terminus derived from an extracellular loop of the bacterial outer membrane protein; and a passenger polypeptide linked or fused directly to the non-native N-terminus or the non-native C-terminus of the bacterial outer membrane protein, wherein the passenger polypeptide is displayed on the outer surface of the host cell and at least one of the non-native N-terminus and the non-native C-terminus is positioned extracellularly. - View Dependent Claims (19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32)
Specification