Methods for producing members of specific binding pairs

• US 6,172,197 B1
• Filed: 06/07/1995
• Issued: 01/09/2001
• Est. Priority Date: 05/15/1992
• Status: Expired due to Term

• Current Assignee: UK Research and Innovation
• Original Assignee: Cambridge Antibody Technology Ltd (Astrazeneca PLC), Medical Research Council (Government of United Kingdom)
• Inventors: 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, Chiswell, David John, Clackson, Timothy Piers, McCafferty, John, Holliger, Kaspar Philipp
• Primary Examiner(s): MacMillan, Keith D.
• Assistant Examiner(s): PONNALURI, PADMASHRI
• Application Number:
  US08/484,893
• Time in Patent Office: 2,043 Days
• Field of Search: 435/5, 435/252.3, 435/6, 435/69.6, 435/7.1, 435/69.7, 435/9.1, 435/69.1, 435/69.8, 435/205.1, 435/320.1, 435/DIG.22, 435/DIG.23, 435/DIG.34, 435/DIG.35, 435/DIG.24, 530/387.1, 530/387.3, 530/412, 530/867, 536/23.1, 536/23.53, 536/23.4
• US Class Current: 530/387.3
• CPC Class Codes: C07K 16/18 against material from animals or humans
• 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 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: Y10S 530/867 involving immunoglobulin or antibody produced via recombinant dna technology

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 library of filamentous bacteriophage particles displaying on their surface as a fusion with a gene III coat protein surface component a genetically diverse population of specific binding pair members in functional form comprising a binding domain for complementary binding specific binding pair members, said specific binding pair members encoded by nucleic acid derived from a natural repertoire of nucleic acids encoding said genetically diverse population of specific binding pair members, the particles each containing a phagemid genome which is plasmid nucleic acid containing a single stranded phage replication origin and a nucleotide sequence encoding said fusion, and the particle having a coat partially derived from a helper phage and partly from said fusion.

2. A library according to claim 1 wherein the specific binding pair members comprise a binding domain of an immunoglobulin.

3. A library according to claim 2 wherein the specific binding pair members are scFv molecules.

4. A library according to claim 2 wherein the specific binding pair members comprise Fab molecules.

5. The library of claims 2, 3, or 4 wherein the natural repertoire of specific binding pair members is encoded by nucleic acid derived from an animal unimmunized against the complementary specific binding pair member.

6. The library of claims 2, 3, or 4 wherein the natural repertoire of specific binding pair members is encoded by nucleic acid derived from an animal immunized against the complementary specific binding pair member.