Improvements to ribosome display

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/10 (2006.01) C07K 16/00 (2006.01) C07K 16/44 (2006.01) C12N 7/00 (2006.01) C12N 7/04 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2402452

The use in a ribosome display system for a selection of a specific binding pair member (e.g. antibody molecule) able to bind a complementary specific binding pair member (e.g. antigen) of encapsidating specific binding member/ribosome complexes ina viral coat, optionally in combination with incorporation of an Midvariant RNA template and optionally one or more other improvements selected from : glycine-serine tether, protein disulphide isomerase, protein disulphide isomerase in combination with oxidised and reduced glutathione at a ratio of between 1:1 and 10:1, addition of oxidised and reduced glutathione at a ratio of between 1:1 and 10:1 after 30 minutes of in vitro translation; blocking with heparin during selection.

L'invention concerne l'utilisation, dans un système d'exposition du ribosome servant à la sélection d'un élément d'une paire de liaison spécifique (p. ex. une molécule d'anticorps) pouvant se lier à un élément d'une paire de liaison spécifique complémentaire (p. ex. un antigène), de complexes d'encapasidation d'élément de liaison spécifique et du ribosome dans une enveloppe virale, éventuellement en combinaison avec l'incorporation d'une matrice d'ARN Midvariant, et éventuellement une ou plusieurs autres améliorations choisies entre: la liaison glycine-sérine, le disulfure-isomérase protéique, le disulfure-isomérase protéique combiné au glutathione oxydé et réduit selon un rapport compris entre 1:1 et 10:1, l'adjonction de glutathione oxydé et réduit selon un rapport compris entre 1:1 et 10:1 après 30 minutes de traduction in vitro, le tout assorti du blocage de l'héparine pendant la sélection.

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