Methods for producing members of specific binding pairs

• US 6,806,079 B1
• Filed: 11/28/2000
• Issued: 10/19/2004
• Est. Priority Date: 07/10/1990
• Status: Expired due to Fees

• Current Assignee: UK Research and Innovation
• Original Assignee: Medical Research Council (Government of United Kingdom), Cambridge Silicon Radio Limited (Qualcomm, Inc.)
• Inventors: Chiswell, David John, Clackson, Timothy Piers, Bonnert, Timothy Peter, Jackson, Ronald Henry, Marks, James David, Griffiths, Andrew David, Winter, Gregory Paul, Hoogenboom, Henricus Renerus Jacobus Mattheus, Pope, Anthony Richard, Johnson, Kevin Stuart, McCafferty, John, Holliger, Kaspar Philipp
• Primary Examiner(s): Ponnaluri, Padmashri
• Application Number:
  US09/726,219
• Time in Patent Office: 1,421 Days
• Field of Search: 435/5, 435/6, 435/69.1, 435/69.7, 435/320.1, 435/DIG.1, 435/DIG.2, 435/DIG.34, 435/DIG.37, 435/DIG.47, 530/387.1, 530/387.3, 536/23.1, 536/23.4
• US Class Current: 435/320.1
• CPC Class Codes: B01J 2219/00718 Type of compounds synthesised
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/34 against blood group antigens
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 16/44 against material not provided for elsewhere , e.g. haptens, metals, DNA, RNA, amino acids
• CPC Class Codes: C07K 16/462 Igs containing a variable region (Fv) from one specie and a constant region (Fc) from another
• CPC Class Codes: C07K 16/468 Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
• CPC Class Codes: C07K 19/00 Hybrid peptides , i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C07K 2319/41 containing a Myc-tag
• CPC Class Codes: C07K 2319/735 containing a domain for self-assembly, e.g. a viral coat protein (includes phage display)
• CPC Class Codes: C12N 15/1037 Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/73 Expression systems using phage (lambda) regulatory sequences
• 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
• CPC Class Codes: G01N 33/53 : Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/6857 : Antibody fragments

Abstract

A member of a specific binding pair (sbp) is identified by expressing DNA encoding a genetically diverse population of such sbp members in recombinant host cells in which the sbp members are displayed in functional form at the surface of a secreted recombinant genetic display package (rgdp) containing DNA encoding the sbp member or a polypeptide component thereof, by virtue of the sbp member or a polypeptide component thereof being expressed as a fusion with a capsid component of the rgdp. The displayed sbps may be selected by affinity with a complementary sbp member, and the DNA recovered from selected rgdps for expression of the selected sbp members. Antibody sbp members may be thus obtained, with the different chains thereof expressed, one fused to the capsid component and the other in free form for association with the fusion partner polypeptide. A phagemid may be used as an expression vector, with said capsid fusion helping to package the phagemid DNA. Using this method libraries of DNA encoding respective chains of such multimeric sbp members may be combined, thereby obtaining a much greater genetic diversity in the sbp members than could easily be obtained by conventional methods.

Claims

1. A filamentous bacteriophage particle displaying on its surface a binding molecule comprising a binding domain able to bind target epitope or antigen, which binding molecule is selected from the group consisting of a Fab antibody molecule, a single chain Fv antibody molecule and an antibody VH domain, the particle containing a phagemid genome comprising nucleic acid encoding the binding molecule, or a polypeptide chain component of the binding molecule.

2. A filamentous bacteriophage particle according to claim 1 wherein said binding molecule is a scFv antibody molecule.

3. A filamentous bacteriophage particle according to claim 1 wherein said binding molecule is a Fab antibody molecule.

4. A filamentous bacteriophage particle according to claim 1 wherein said binding molecule is an antibody VH domain.

5. A filamentous bacteriophage particle according to any one of claims 1 to 4, which is in a population of filamentous bacteriophage particles displaying a population of said binding molecules having a range of binding specificities.