Polypeptide Display Libraries and Methods of Making and Using Thereof
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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.
3 Citations
56 Claims
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1-20. -20. (canceled)
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21. A composition comprising:
a fusion protein, the fusion protein comprising a bacterial outer membrane protein, 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 (22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39)
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40. 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 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 (41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56)
Specification