Production of chimeric antibodies - a combinatorial approach
First Claim
1. A method of making antibody polypeptide dimers specific for an antigen of interest, having the following steps:
- (i) providing nucleic acid expression vectors which are packaged using a component of a replicable genetic display package;
(ii) combining (a) a genetically diverse repertoire of nucleic acid sequences which each encode a first component part of an antigen-binding site of a human antibody with (b) nucleic acid which encodes a unique or genetically diverse population of a second component part of an antigen-binding site of a non-human antibody known to bind said antigen of interest, to form a library of nucleic acid sequences on said expression vectors encoding antibody polypeptide dimers, which dimers each consist of a first polypeptide chain component and a second polypeptide chain component, in combination forming an antigen-binding site of an antibody polypeptide dimer, wherein said library contains nucleic acid encoding an antibody polypeptide dimer specific for an antigen of interest;
(iii) expressing said library from said vectors in recombinant host organism cells, each of the said first polypeptide chain components being expressed as a fusion with a component of a replicable genetic display package which thereby displays said first polypeptide chain component at the surface of replicable genetic display packages;
(iv) selecting from said expressed library by binding with antigen a unique or restricted population of said antibody polypeptide dimers which have binding specificity for said antigen of interest, each selected antibody polypeptide dimer being associated in its respective replicable genetic display package with nucleic acid encoding said first component part of the antigen-binding site thereof.
4 Assignments
0 Petitions
Accused Products
Abstract
Methods are disclosed which may be used for the production of antibodies, or antibody fragments, which have the same binding specificity as a parent antibody but which have increased human characteristics. Humanized antibodies may be obtained by chain shuffling, perhaps using phage display technology. In one embodiment, a polypeptide comprising a heavy or light chain variable domain of a non-human antibody specific for an antigen of interest is combined with a repertoire of human complementary (light or heavy) chain variable domains. Hybrid pairings which are specific for the antigen of interest are selected. Human chains from the selected pairings may then be combined with a repertoire of human complementary variable domains (heavy or light) and humanized antibody polypeptide dimers can then be selected for binding specificity for antigen. The methods may be combined with CDR-imprinting. In another embodiment, component part of an antigen-binding site of a no-human antibody known to bind a particular antigen is combined with a repertoire of component parts of an antigen-binding site of human antibody, forming a library of antibody polypeptide dimers with antigen-binding sites. Hybrids selected from this library may be used in a second humanizing shuffling step, or may already be of sufficient human character to be of value, perhaps after some modification to increase human character still further.
1633 Citations
26 Claims
-
1. A method of making antibody polypeptide dimers specific for an antigen of interest, having the following steps:
-
(i) providing nucleic acid expression vectors which are packaged using a component of a replicable genetic display package; (ii) combining (a) a genetically diverse repertoire of nucleic acid sequences which each encode a first component part of an antigen-binding site of a human antibody with (b) nucleic acid which encodes a unique or genetically diverse population of a second component part of an antigen-binding site of a non-human antibody known to bind said antigen of interest, to form a library of nucleic acid sequences on said expression vectors encoding antibody polypeptide dimers, which dimers each consist of a first polypeptide chain component and a second polypeptide chain component, in combination forming an antigen-binding site of an antibody polypeptide dimer, wherein said library contains nucleic acid encoding an antibody polypeptide dimer specific for an antigen of interest; (iii) expressing said library from said vectors in recombinant host organism cells, each of the said first polypeptide chain components being expressed as a fusion with a component of a replicable genetic display package which thereby displays said first polypeptide chain component at the surface of replicable genetic display packages; (iv) selecting from said expressed library by binding with antigen a unique or restricted population of said antibody polypeptide dimers which have binding specificity for said antigen of interest, each selected antibody polypeptide dimer being associated in its respective replicable genetic display package with nucleic acid encoding said first component part of the antigen-binding site thereof. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 23)
-
-
10. A method of making human antibody polypeptide dimers specific for an antigen of interest, comprising:
-
(i) combining a diverse population of polypeptides each comprising a variable domain of a first polypeptide chain of a human antibody (population A) with a unique or restricted population of polypeptides each comprising a variable domain of a second polypeptide chain of a non-human antibody specific for said antigen (population B), thereby forming a library of antibody polypeptide dimers each consisting of a polypeptide which comprises a variable domain of a first polypeptide chain of a human antibody and a polypeptide chain of a non-human antibody, wherein said library contains an antibody polypeptide dimer specific for an antigen of interest; (ii) selecting from said library a unique or restricted population of said antibody polypeptide dimers which have binding specificity for said antigen (population c); (iii) combining a unique or restricted population of human polypeptides derived from polypeptide dimers selected in step (ii) each comprising a human first polypeptide chain (population D) with a diverse population of polypeptides each comprising a variable domain of a second polypeptide chain of a human antibody (population E), thereby forming a library of human antibody polypeptide chain dimers from which a unique or restricted population chain dimers from which a unique or restricted population of human antibody polypeptide dimers specific for said antigen (population F) are selectable. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 24, 25, 26)
-
Specification