Methods for display of heterodimeric proteins on filamentous phage using pVII and pIX, compositions, vectors and combinatorial libraries

• US 6,472,147 B1
• Filed: 05/25/1999
• Issued: 10/29/2002
• Est. Priority Date: 05/25/1999
• Status: Expired due to Term

• Current Assignee: The Scripps Research Institute
• Original Assignee: The Scripps Research Institute
• Inventors: Gao, Changshou, Lerner, Richard A., Janda, Kim D., Wirsching, Peter
• Primary Examiner(s): Ponnaluri, Padmashri
• Application Number:
  US09/318,786
• Time in Patent Office: 1,253 Days
• Field of Search: 435/5, 435/6, 435/4, 435/7.1, 435/DIG.1, 435/DIG.23, 435/DIG.24, 435/69.1, 435/69.7, 435/320.1, 424/162.1, 424/93.6, 530/300, 530/350, 530/387.1
• US Class Current: 506/14
• CPC Class Codes: B01J 2219/0072 Organic compounds
• 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 describes the display of exogenous polypeptides on filamentous phage using a fusion between the exogenous polypeptide and phage pVII or pIX proteins. In particular, phage particles and phagemid vectors are described for expression and display of heterodimeric proteins such a antibody Fv heterodimers in combinatorial libraries, and uses thereof.

Claims

1. A filamentous phage encapsulating a genome encoding a fusion polypeptide and having said fusion polypeptide on a surface of said phage, wherein said polypeptide comprises an exogenous polypeptide fused to an amino terminus of a filamentous phage pVII or pIX protein.

2. The filamentous phage of claim 1 wherein said exogenous polypeptide is selected from the group consisting of an immunoglobulin heavy chain variable domain (V_H ), an immunoglobulin light chain variable domain (V_L ), a synthetic polypeptide and a single chain antibody (scFv).

3. The filamentous phage of claim 1 wherein said genome is pCGMT or pCGMT-1b having a nucleotide sequence shown in SEQ ID NO 19 or 20, respectively.

4. The filamentous phage of claim 2 wherein said scFv is scFv-21H3-pIX.

5. A library of filamentous phage particles wherein each phage particle is according to claim 1.