Adaptor-directed helper systems

• US 7,910,350 B2
• Filed: 12/19/2005
• Issued: 03/22/2011
• Est. Priority Date: 11/02/2001
• Status: Active Grant

• Current Assignee: Abmaxis, Inc. (Merck & Co., Inc.)
• Original Assignee: Abmaxis, Inc. (Merck & Co., Inc.)
• Inventors: Zhong, Pingyu, Wang, Caili, Wang, Xinwei
• Primary Examiner(s): Ketter; James S
• Application Number:
  US11/313,270
• Publication Number:
  US 20060281181A1
• Time in Patent Office: 1,919 Days
• Field of Search: None
• US Class Current: 435/235.1
• 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 present invention provides adapter-directed display systems for expressing exogenous polypeptide within a host cell and/or displaying the exogenous polypeptide on the outer surface of a genetic package. This subject systems are particularly useful for displaying a genetically diverse repertoire of monomeric and multimeric polypeptides. The invention also provides both expression and helper vectors and kits containing components of the subject display systems. Also provided are genetic packages displaying the exogenous polypeptides of particular interest. Further provided by the invention are methods of using the subject display systems.

Claims

1. A phage helper vector comprising a nucleic acid sequence which encodes all of the outer-surface sequences required to package a phage genome wherein at least one of the outer-surface sequences is a recombinant protein comprising a coat protein fused in-frame to an adapter sequence.

2. The helper vector of claim 1, wherein the the phage is filamentous phage and the recombinant protein is a coat protein encoded by a gene selected from the group consisting of gene III, gene VI, gene VII, gene VIII, and gene IX.

3. The helper vector of claim 2, wherein the recombinant protein is pIII.

4. The helper vector of claim 2, wherein the adapter sequence is capable of pairwise interaction with a corresponding adapter to form a stable complex.

5. The helper vector of claim 4, wherein the adapter is derived from a heterodimeric receptor and adopts a coiled coil structure.

6. The helper vector of claim 4, wherein the adapter is derived from a GABA_B receptor.

7. The helper vector of claim 4, wherein upon infection of a host cell carrying a phagemid vector which comprises a library of exogenous polypeptide sequences fused in frame to an adapter that is capable of pairwise interaction with the adapter fused in frame to the recombinant coat protein, the helper vector functions to cause the exogenous sequences to be displayed on the surface of packaged phage particles.

8. The helper vector of claim 6, wherein the adapter sequence is derived from GABA_B receptor 2.

9. The helper vector of claim 7, wherein the adapter sequences further comprises the spacer sequence ValGlyGlyCys.

11. The helper vector of claim 1, wherein the vector optionally comprises one or more copies of the outer-surface coat protein selected for use as the recombinant fusion protein.

10. A phage helper vector selected from GM-UltraHelper phage vector, CM-UltraHelper phage vector, and GMCT-UltraHelper phage vector.