IP Library Granted Patent US 9,062,312
Granted Patent B2
US 9,062,312 · App. 13/528,201 · Granted Jun 23, 2015

Fusion peptides comprising multi-functional peptidic solubility tags for efficient production, processing and surface applications

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Quick Facts
Patent No.
US 9,062,312
App. No.
13/528,201
Granted
Jun 23, 2015
Kind
B2
Abstract

Compositions and methods using fusion peptides comprising multi-functional solubility tags are provided. The multi-functional peptidic solubility tags facilitates more efficient fusion peptide production, easier downstream processing of the fusion peptide, and may be used to provide functional surface properties when coupled to a target material.

Claims (17)

1. A method comprising:

a) providing a microbial host cell comprising a heterologous nucleic acid molecule encoding an insoluble fusion peptide comprising at least one first portion and at least one second portion, wherein said first portion comprises a multi-functional solubility tag and said second portion comprises a peptide of interest; wherein the multi-functional solubility tag comprises at least two core motifs independently selected from the group consisting of SEQ ID NOs: 80-104;

b) growing the microbial host cell under conditions whereby the insoluble fusion peptide is produced within the microbial host cell in the form of at least one inclusion body;

c) recovering the insoluble fusion peptide from the microbial host cell;

d) subjecting the insoluble fusion peptide to an aqueous medium having a pH at which the insoluble fusion peptide becomes a soluble fusion peptide;

e) contacting the soluble fusion peptide with a first target material having a surface whereby the fusion peptide non-covalently associates, with a KD or MBs0 value of less than or equal to about 10-5 M, with the surface of the first target material; wherein the association between the fusion peptide and the first target material is dependent upon, or enhanced by, the presence of the multi-functional solubility tag; and

(f) altering the pH in step (d) to promote or enhance deposition of the soluble fusion peptide on the surface of the first target material.

2. The method of claim 1 wherein pH is increased.

3. The method of claim 1 wherein pH is decreased.

4. The method of claim 1 wherein pH is altered by at least one pH unit.

5. The method of claim 1 wherein the second portion of the fusion peptide is an antimicrobial agent.

6. The method of claim 1 wherein the second portion of the fusion peptide has an affinity for a second target material; wherein the first target material and the second target material are different.

7. The method of claim 1 wherein the first target material is a particle having an average particle size ranging from 100 nm to 10 μas measured by a light scattering method.

8. The method of claim 1 wherein the first target material is selected from the group consisting of a pigment, a body tissue, a cell surface receptor, hair, skin, nail, teeth, oral cavity tissues, plant tissue, a polymer, polystyrene, polyethylene, polypropylene, polytetrafluoroethylene, polyester, polyvinyl chloride, poly (methyl methacrylate), polyethersulfone, polyimide, polyamide, aramids, poly para-phenylene terephthalamide, poly meta-phenylene terephthalamide, poly urethanes, polysaccharides, cellulose, starch, chitin, minerals, silica, silicates, micas, titanium dioxide, alumina, iron oxides, clays, metals, gold, silver, carbon, carbon black, graphite, and carbon nanotubes.

9. The method of claim 6 wherein the second target material is a keratin-containing material selected from the group consisting of skin, nail, and hair.

10. The method of claim 1 wherein the multi-functional solubility tag comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 4, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, and 77.

11. The method of claim 1 where the multi-functional solubility tag comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 37, 40, 42, 43, 44, 45, 46, 47, 49, 50, 51, 62, 63, 64, 65, 66 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, and 77.

Assignments (4)
CORRECTIVE ASSIGNMENT TO CORRECT THE DESIGNATION OF ASSIGNEE AS A DELAWARE CORPORATION TO A DELAWARE LIMITED LIABILITY COMPANY PREVIOUSLY RECORDED ON REEL 049880 FRAME 0001. ASSIGNOR(S) HEREBY CONFIRMS THE ASSIGNMENT. Recorded Aug 28, 2019
From: E. I. DU PONT DE NEMOURS AND COMPANY
To: DUPONT US HOLDING, LLC
Reel/Frame 051502/0610 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jul 26, 2019
From: E. I. DU PONT DE NEMOURS AND COMPANY
To: DUPONT US HOLDING, LLC
Reel/Frame 049880/0001 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 1, 2012
From: ROUVIERE, PIERRE E.; GRUBER, TANJA MARIA; FOSSER, KARI A.; FAHNESTOCK, STEPHEN R.
To: E. I. DU PONT DE NEMOURS AND COMPANY
Reel/Frame 028889/0464 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 28, 2012
From: ROUVIERE, PIERRE E.; GRUBER, TANJA MARIA; FOSSER, KARI A.; FAHNESTOCK, STEPHEN R.
To: E. I. DU PONT DE NEMOURS AND COMPANY
Reel/Frame 028865/0042 →