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Contenido archivado el 2022-12-23

Relationship between haptenic structure and properties of the antibody binding sites for antibodies obtained from hybridomas and from phage display libraries

Objetivo



Antibodies elicited against a transition state analogue of a reaction are able to catalyse that reaction. This means that catalytic antibodies, in contrast to enzymes, can in principle be prepared for any desired reaction. Binding domains of catalytic antibodies can be cloned and expressed in E.coli providing an unlimited and cheap source of potentially very efficient and stable biocatalysts. In a novel approach a library of between 106-108 different antibody fragments are displayed on a phage surface. Antibodies with the desired binding properties can easily be selected from the total population. These developments facilitate the application of antibodies as, e.g. industrial catalysts.

For proper hapten design, to elicit efficient antibodies for a desired reaction it is important to investigate the relationship between haptenic structure and the properties of the antibody combining site.

The research will involve hapten synthesis and the preparation of monoclonal antibodies for a decarboxylation reaction. Furthermore, a number of haptens of related structures will be synthesised to elicit at a later stage antibodies for different types of reaction, e.g. enantioselective hydrogenation of carbonyl in diheterylketones, reduction of carbon-carbon double bonds in a,b-unsaturated ketones, ester and amide hydrolysis and cyanohydrination. Substrates and haptens will be synthesised in Kiev and in Cambridge. In Moscow monoclonal antibodies against these haptens will be prepared using hybridoma technology. In Wageningen a phage display library will be screened for hapten binding.

The antibodies/antibody fragments will be tested for catalytic activity and the most promising ones will be further characterised by kinetic and thermodynamic studies. Selected antibodies obtained via hybridoma technology will be cloned and sequenced to allow comparison with antibody fragments obtained from the library at the molecular level.

Convocatoria de propuestas

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Régimen de financiación

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Coordinador

University of Cambridge
Aportación de la UE
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Dirección
Lensfield Road
CB2 1EW Cambridge
Reino Unido

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Participantes (3)