Major coat protein variants for C-terminal and bi-terminal display

• US 20060292554A1
• Filed: 05/17/2005
• Published: 12/28/2006
• Est. Priority Date: 05/18/2004
• Status: Abandoned Application

• Current Assignee: Genentech, Inc. (Roche Holding AG)
• Original Assignee: Genentech, Inc. (Roche Holding AG)
• Inventors: Held, Heike A., Sidhu, Sachdev S.
• Application Number:
  US11/131,109
• Publication Number:
  US 20060292554A1
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: C12N 15/1037 Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
• CPC Class Codes: C40B 40/02 Libraries contained in or displayed by microorganisms, e.g. bacteria or animal cells; Libraries contained in or displayed by vectors, e.g. plasmids; Libraries containing only microorganisms or vectors

Abstract

The invention provides compositions and methods for enhanced display of heterologous proteins on viral particles.

Claims

1. A fusion protein comprising a heterologous polypeptide fused to a major coat protein of a virus, wherein the major coat protein is a variant of a wild type major coat protein of the virus and is capable of C-terminal display of the heterologous polypeptide at a display level more than 30 times that of a corresponding coat protein comprising a wild type sequence.

4. A replicable expression vector comprising a gene fusion, wherein the gene fusion encodes the fusion protein of claim 1, 2 or 3.

5. A library comprising a plurality of the replicable expression vectors of claim 4, the expression vectors comprising a plurality of different gene fusions encoding a plurality of fusion proteins.

6. A host cell comprising the vector of claim 4.

7. A virus displaying the fusion protein of claim 1 or 2 on the surface thereof.

8. A library of virus, comprising a plurality of the virus of claim 7 displaying a plurality of different fusion proteins on the surface thereof.

9. A method, comprising:

constructing a library of phage or phagemid particles displaying a plurality of the fusion protein of claim 1 or 2;

contacting the phage or phagemid particles with a target molecule or substance; and

separating particles having a desired selection characteristic from those that do not.

2. A fusion protein comprising a major coat protein of a virus fused on its N-terminus to a first heterologous polypeptide and on its C-terminus to a second heterologous polypeptide, wherein the major coat protein is a variant of a wild type major coat protein of the virus.

3. The fusion protein of claim 2 wherein the variant coat protein is capable of C-terminal display of the heterologous polypeptide at a display level more than 30 times that of a corresponding coat protein comprising a wild type sequence.