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.
5 Citations
52 Claims
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1-20. -20. (canceled)
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21. A method for identifying a target in a sample, the method comprising:
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(a) contacting the sample with a fusion protein, wherein the fusion protein comprises an outer membrane protein and a passenger polypeptide, (i) wherein a native N-terminus and a native C-terminus of the outer membrane protein are joined by a linker, (ii) wherein the outer membrane protein comprises a non-native N-terminus and a non-native C-terminus derived from an extracellular loop of the outer membrane protein, and (iii) wherein the passenger polypeptide is linked or fused to the non-native N-terminus or the non-native C-terminus of the outer membrane protein; and (b) detecting a specific interaction between the passenger polypeptide and the target, wherein the presence of the specific interaction is indicative of the presence of the target in the sample. - View Dependent Claims (22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38)
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39. A method for identifying a target in a sample, the method comprising:
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(a) contacting the sample with a plurality of fusion proteins, wherein each fusion protein of the plurality comprises an outer membrane protein and a passenger polypeptide, (i) wherein a native N-terminus and a native C-terminus of the outer membrane protein are joined by a linker, (ii) wherein the outer membrane protein comprises a non-native N-terminus and a non-native C-terminus derived from an extracellular loop of the outer membrane protein, and (iii) wherein the passenger polypeptide is linked or fused to the non-native N-terminus or the non-native C-terminus of the outer membrane protein; and (b) detecting a specific interaction between at least one of the passenger polypeptides of the plurality of fusion proteins and the target, wherein the presence of the specific interaction is indicative of the presence of the target in the sample. - View Dependent Claims (40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52)
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Specification