Methods and reagents for isolating biologically active peptides
OwnerJeno Gyuris, Aaron J. Morrisinventors
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Abstract
One aspect of the present invention is the synthesis of a binary method that combines variegated peptide display libraries, e.g., in a “display mode”, with soluble secreted peptide libraries, e.g., in a “secretion mode”, to yield a method for the efficient isolation of peptides having a desired biological activity.
Claims79
A method for generating a peptide with a selected biological activity, comprising the steps of:
- (i) providing a peptide display library comprising a variegated population of test peptides expressed on the surface of a population of display packages;
- (ii) in a display mode, isolating, from the peptide display libary, a sub-population of display packages enriched for test peptides which have a desired binding specificity and/or affinity for a cell or a component thereof;
- (iii) in a secretion mode, simultaneously expressing the enriched test peptide sub-population under conditions wherein the test peptides are secreted and are free of the display packages;
- and
- (iv) assessing the ability of the secreted test peptides to regulate a biological process in a target cell.
An peptide display library enriched for test peptides having a desired binding specificity and/or affinity for a cell or a component thereof and which regulate a biological process in a target cell.
A vector comprising a chimeric gene for a chimeric protein, which chimeric gene comprises (i) a coding sequence for a test peptide, (ii) a coding sequence for a surface protein of a display package, and (iii) RNA splice sites flanking the coding sequence for the surface protein, wherein,
- in a display mode, the chimeric gene is expressed as a fusion protein including the test peptide and the surface protein such that the test peptide can be displayed on the surface of a population of display packages,
- whereas in the secretion mode, the test peptide is expressed without the surface protein as a result of the coding sequence for the surface protein being removed by RNA splicing.
A vector library, each vector comprising a chimeric gene for a chimeric protein, which chimeric gene comprises (i) a coding sequence for a test peptide, (ii) a coding sequence for a surface protein of a display package, and (iii) RNA splice sites flanking the coding sequence for the surface protein, wherein,
- in a display mode, the chimeric gene is expressed as fusion protein including the test peptide and the surface protein such that the test peptide can be displayed on the surface of a population of display packages,
- whereas in the secretion mode, the test peptide is expressed without the surface protein as a result of the coding sequence for the surface protein being removed by RNA splicing, the vector library collectively encodes a variegated population of test peptides.
A method for generating a peptide with a selected antimicrobial activity, comprising the steps of:
- (i) providing a recombinant host cell population which expresses a soluble peptide library comprising a variegated population of test peptides;
- (ii) culturing the host cells with a target microorganism under conditions wherein the peptide library is secreted and diffuses to the target microorganism;
- and
- (iii) selected host cells expressing test peptides that inhibit growth of the target microorganism.
A construct as shown in FIG. 1, 2 or 3.
Dependencies are inferred from the claim text.