AFFINITY LABELLING OF ANTIBODIES WITH ARYL NITRENE AS REACTIVE GROUP

THE covalent binding of a haptene to its specific antibody with a view to identifying the section of peptide chains which form the combining site has been pioneered by Singer et al.1,2, who named this approach affinity labelling. Antibodies were prepared in rabbits against several aromatic haptenes...

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Main Authors: Fleet, G, Porter, R, Knowles, J
Format: Journal article
Language:English
Published: 1969
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author Fleet, G
Porter, R
Knowles, J
author_facet Fleet, G
Porter, R
Knowles, J
author_sort Fleet, G
collection OXFORD
description THE covalent binding of a haptene to its specific antibody with a view to identifying the section of peptide chains which form the combining site has been pioneered by Singer et al.1,2, who named this approach affinity labelling. Antibodies were prepared in rabbits against several aromatic haptenes such as benzene arsonate, and affinity labelling was performed by reaction with p-(arsonic acid)-benzene diazonium fluoroborate through the reactive diazonium group. The reaction with specific antibody was considerably more rapid than that with inert IgG because of the concentration of reagent in the combining site. Subsequent separation of heavy and light chains followed by enzymic digestion led to the isolation of one predominant dipeptide (Val-Tyr) from the light chain and another (Thr-Tyr) from the heavy chain; both of these were substituted on the tyrosine residue. No sequence data were available on the peptide chains of rabbit IgG, but from comparison with the known sequences of the light chains of human and mouse IgG it was suggested that the tyrosine residue 86 in the light chain may have been the one labelled. © 1969 Nature Publishing Group.
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spelling oxford-uuid:bab76b11-905d-4141-8e16-d27c03d9f3072022-03-27T05:11:46ZAFFINITY LABELLING OF ANTIBODIES WITH ARYL NITRENE AS REACTIVE GROUPJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:bab76b11-905d-4141-8e16-d27c03d9f307EnglishSymplectic Elements at Oxford1969Fleet, GPorter, RKnowles, JTHE covalent binding of a haptene to its specific antibody with a view to identifying the section of peptide chains which form the combining site has been pioneered by Singer et al.1,2, who named this approach affinity labelling. Antibodies were prepared in rabbits against several aromatic haptenes such as benzene arsonate, and affinity labelling was performed by reaction with p-(arsonic acid)-benzene diazonium fluoroborate through the reactive diazonium group. The reaction with specific antibody was considerably more rapid than that with inert IgG because of the concentration of reagent in the combining site. Subsequent separation of heavy and light chains followed by enzymic digestion led to the isolation of one predominant dipeptide (Val-Tyr) from the light chain and another (Thr-Tyr) from the heavy chain; both of these were substituted on the tyrosine residue. No sequence data were available on the peptide chains of rabbit IgG, but from comparison with the known sequences of the light chains of human and mouse IgG it was suggested that the tyrosine residue 86 in the light chain may have been the one labelled. © 1969 Nature Publishing Group.
spellingShingle Fleet, G
Porter, R
Knowles, J
AFFINITY LABELLING OF ANTIBODIES WITH ARYL NITRENE AS REACTIVE GROUP
title AFFINITY LABELLING OF ANTIBODIES WITH ARYL NITRENE AS REACTIVE GROUP
title_full AFFINITY LABELLING OF ANTIBODIES WITH ARYL NITRENE AS REACTIVE GROUP
title_fullStr AFFINITY LABELLING OF ANTIBODIES WITH ARYL NITRENE AS REACTIVE GROUP
title_full_unstemmed AFFINITY LABELLING OF ANTIBODIES WITH ARYL NITRENE AS REACTIVE GROUP
title_short AFFINITY LABELLING OF ANTIBODIES WITH ARYL NITRENE AS REACTIVE GROUP
title_sort affinity labelling of antibodies with aryl nitrene as reactive group
work_keys_str_mv AT fleetg affinitylabellingofantibodieswitharylnitreneasreactivegroup
AT porterr affinitylabellingofantibodieswitharylnitreneasreactivegroup
AT knowlesj affinitylabellingofantibodieswitharylnitreneasreactivegroup