Assay methods and materials for distinguishing analytes and...

G - Physics – 01 – N

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G01N 33/53 (2006.01) G01N 33/543 (2006.01)

Patent

CA 2537494

The invention relates to assays in which complexes between an analyte and a binding agent specific for the analyte are detected by a reporter molecule, which is capable of binding to the binding agent when the binding agent is complexed with analyte but not when the binding agent is complexed with an analyte analogue. The invention provides a method of modifying such a binding agent in order to create a tailored site where the reporter molecule may bind to the binding agent. This provides control over the location and nature of the reporter binding site. Antibodies, including bispecific antibodies, are preferred binding agents. The invention also provides novel methods for labelling antibodies close to their antigen binding site using bi- and tri- functional reagents. Modified binding agents produced by the methods described are also provided.

L'invention concerne des analyses dans lesquelles des complexes entre un analyte et un agent de liaison spécifique de l'analyte sont détectés par une molécule rapporteur, laquelle peut se lier à l'agent de liaison lorsque l'agent de liaison est complexé avec un analyte mais pas lorsque l'agent de liaison est complexé avec un analogue d'analyte. L'invention concerne une méthode permettant de modifier un tel agent de liaison de manière à créer un site adapté où la molécule rapporteur peut se lier à l'agent de liaison, ce qui permet de contrôler l'emplacement et la nature du site de liaison du rapporteur. Les anticorps, notamment les anticorps bispécifiques, constituent des agents de liaison préférés. L'invention concerne également de nouvelles méthodes de marquage d'anticorps à proximité de leur site de liaison aux antigènes à l'aide de réactifs bifonctionnels et trifonctionnels. L'invention concerne en outre des agents de liaison modifiés produits selon lesdites méthodes.

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