Recombinant altered antibodies and methods of making altered antibodies
AssigneeMedical Research Council
InventorGregory P. Winter
Life of this patent
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Abstract
An altered antibody is produced by replacing the complementarity determining regions (CDRs) of a variable region of an immunoglobulin (Ig) with the CDRs from an Ig of different specificity, using recombinant DNA techniques. The gene coding sequences for producing the altered antibody may be produced by site-directed mutagenesis using long oligonucleotides.
Claims23
An altered antibody or antigen-binding fragment thereof, wherein a variable domain of the antibody or antigen-binding fragment has the framework regions of a first immunoglobulin heavy or light chain variable domain and the complementarity determining regions of a second immunoglobulin heavy or light chain variable domain, wherein said second immunoglobulin heavy or light chain variable domain is different from said first immunoglobulin heavy or light chain variable domain in antigen binding specificity, antigen binding affinity, species, class or subclass.
A method of producing an altered antibody comprising the steps:
- (a) preparing a eukaryotic expression vector comprising a promoter operably linked to a DNA sequence which encodes an immunoglobulin heavy or light chain, wherein the variable domain of said immunoglobulin heavy or light chain comprises the framework regions of a first immunoglobulin heavy or light chain variable domain and the complementarity determining regions of a second heavy or light chain variable domain, and wherein said second immunoglobulin heavy or light chain variable domain is different from said first immunoglobulin heavy or light chain variable domain in antigen binding specificity, antigen binding affinity, species, class or subclass;
- (b) transforming a cell line with said expression vector;
- (c) culturing said transformed cell line to produce said altered antibody;
- and,
- (d) recovering said altered antibody.
A method of producing an altered antibody comprising the steps:
- (a) preparing a first eukaryotic expression vector comprising a promoter operably linked to a DNA sequence which encodes an immunoglobulin heavy chain, wherein the variable domain of said immunoglobulin heavy chain comprises the framework regions of a first immunoglobulin heavy chain variable domain and the complementarity determining regions of a second heavy chain variable domain, and wherein said second immunoglobulin heavy chain variable domain is different from said first immunoglobulin heavy chain variable domain in antigen binding specificity, antigen binding affinity, species, class or subclass;
- (b) preparing a second eukaryotic expression vector comprising a promoter operably linked to a DNA sequence which encodes an immunoglobulin light chain, wherein the variable domain of said immunoglobulin light chain comprises the framework regions of a first immunoglobulin light chain variable domain and the complementarity determining regions of a second light chain variable domain, and wherein said second immunoglobulin light chain variable domain is different from said first immunoglobulin light chain variable domain in antigen binding specificity, antigen binding affinity, species, class or subclass;
- (c) transforming a cell line with said first and second expression vectors;
- (d) culturing said transformed cell line to produce said altered antibody;
- and,
- (e) recovering said altered antibody.
A eukaryotic expression vector for expression of an altered antibody comprising a DNA sequence encoding an immunoglobulin heavy or light chain, wherein the variable domain has the framework regions of a first immunoglobulin heavy or light chain variable domain and the complementarity determining regions of a second immunoglobulin heavy or light chain variable domain, and wherein said second immunoglobulin heavy or light chain variable domain is different from said first immunoglobulin heavy or light chain variable domain in antigen binding specificity, antigen binding affinity, species, class or subclass.
Dependencies are inferred from the claim text.