Methods and composition for concentrating secreted...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/81 (2006.01) C12N 9/24 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2582808

Methods and compositions are described that relate to obtaining concentrated preparations of secreted recombinant proteins. These proteins are expressed in the form of fusion proteins with a chitin-binding domain (CBD). The fusion proteins are capable of being concentrated in the presence of chitin. Also described is: a shuttle vector that includes a modified LAC4 promoter; a chitinase-negative host cell; a CBD capable of eluting from chitin under non- denaturing conditions; and sterilized chitin, which can be optionally magnetized for facilitating recovery of recombinant protein.

La présente invention a trait à des procédés et des compositions concernant l'obtention de préparations concentrées de protéines recombinantes sécrétées. Ces protéines sont exprimées sous la forme de protéines hybrides avec un domaine de liaison à la chitine. Les protéines hybrides sont capables d'être concentrées en présence de la chitine. L'invention a également trait à un vecteur navette comportant un promoteur LAC4 modifié, une cellule hôte chitinase-négative, un domaine de liaison à la chitine apte à une élution à partir de la chitine dans des conditions non dénaturantes; et de la chitine stérilisée, qui peut éventuellement être magnétisée pour faciliter la récupération d'une protéine recombinante.

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