Site-specific attachment to recombinant antibodies via introduced surface cysteine residues.
Many diagnostic and therapeutic applications of monoclonal antibodies require the covalent linking of effector or reporter molecules to the immunoglobulin polypeptides. Existing methods generally involve the non-selective modification of amino acid side chains, producing one or more randomly distrib...
| Main Authors: | , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
1990
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| _version_ | 1826264199083327488 |
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| author | Lyons, A King, D Owens, R Yarranton, G Millican, A Whittle, N Adair, JR |
| author_facet | Lyons, A King, D Owens, R Yarranton, G Millican, A Whittle, N Adair, JR |
| author_sort | Lyons, A |
| collection | OXFORD |
| description | Many diagnostic and therapeutic applications of monoclonal antibodies require the covalent linking of effector or reporter molecules to the immunoglobulin polypeptides. Existing methods generally involve the non-selective modification of amino acid side chains, producing one or more randomly distributed attachment sites. This results in heterogeneous labelling of the antibody molecules and often to a decrease in antigen-binding due to the modification of residues close to the antigen-binding site. We report a novel strategy for site-specifically labelling antibodies through surface cysteine residues. Examination of molecular structures was used to identify amino acids of the CH1 domain of the IgG heavy chain that were accessible to solvent but not to larger molecules. Site-directed mutagenesis was used to substitute cysteine residues at these positions in the heavy chain of a mouse/human chimaeric version of the tumour-binding monoclonal antibody, B72.3. Expression of the modified antibody genes in mammalian cells yielded correctly assembled proteins that had thiol groups in pre-determined positions and showed no loss of antigen-binding activity. One of the mutants was used to demonstrate the site-specific attachment of a radio-iodinated ligand to the chimaeric B72.3 antibody. |
| first_indexed | 2024-03-06T20:03:56Z |
| format | Journal article |
| id | oxford-uuid:2846f27c-7775-4052-bd89-590e0869019c |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T20:03:56Z |
| publishDate | 1990 |
| record_format | dspace |
| spelling | oxford-uuid:2846f27c-7775-4052-bd89-590e0869019c2022-03-26T12:11:55ZSite-specific attachment to recombinant antibodies via introduced surface cysteine residues.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:2846f27c-7775-4052-bd89-590e0869019cEnglishSymplectic Elements at Oxford1990Lyons, AKing, DOwens, RYarranton, GMillican, AWhittle, NAdair, JRMany diagnostic and therapeutic applications of monoclonal antibodies require the covalent linking of effector or reporter molecules to the immunoglobulin polypeptides. Existing methods generally involve the non-selective modification of amino acid side chains, producing one or more randomly distributed attachment sites. This results in heterogeneous labelling of the antibody molecules and often to a decrease in antigen-binding due to the modification of residues close to the antigen-binding site. We report a novel strategy for site-specifically labelling antibodies through surface cysteine residues. Examination of molecular structures was used to identify amino acids of the CH1 domain of the IgG heavy chain that were accessible to solvent but not to larger molecules. Site-directed mutagenesis was used to substitute cysteine residues at these positions in the heavy chain of a mouse/human chimaeric version of the tumour-binding monoclonal antibody, B72.3. Expression of the modified antibody genes in mammalian cells yielded correctly assembled proteins that had thiol groups in pre-determined positions and showed no loss of antigen-binding activity. One of the mutants was used to demonstrate the site-specific attachment of a radio-iodinated ligand to the chimaeric B72.3 antibody. |
| spellingShingle | Lyons, A King, D Owens, R Yarranton, G Millican, A Whittle, N Adair, JR Site-specific attachment to recombinant antibodies via introduced surface cysteine residues. |
| title | Site-specific attachment to recombinant antibodies via introduced surface cysteine residues. |
| title_full | Site-specific attachment to recombinant antibodies via introduced surface cysteine residues. |
| title_fullStr | Site-specific attachment to recombinant antibodies via introduced surface cysteine residues. |
| title_full_unstemmed | Site-specific attachment to recombinant antibodies via introduced surface cysteine residues. |
| title_short | Site-specific attachment to recombinant antibodies via introduced surface cysteine residues. |
| title_sort | site specific attachment to recombinant antibodies via introduced surface cysteine residues |
| work_keys_str_mv | AT lyonsa sitespecificattachmenttorecombinantantibodiesviaintroducedsurfacecysteineresidues AT kingd sitespecificattachmenttorecombinantantibodiesviaintroducedsurfacecysteineresidues AT owensr sitespecificattachmenttorecombinantantibodiesviaintroducedsurfacecysteineresidues AT yarrantong sitespecificattachmenttorecombinantantibodiesviaintroducedsurfacecysteineresidues AT millicana sitespecificattachmenttorecombinantantibodiesviaintroducedsurfacecysteineresidues AT whittlen sitespecificattachmenttorecombinantantibodiesviaintroducedsurfacecysteineresidues AT adairjr sitespecificattachmenttorecombinantantibodiesviaintroducedsurfacecysteineresidues |