Magnetically activated cell sorting for production of proteins

C - Chemistry – Metallurgy – 07 – H

Patent

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Details

C07H 21/04 (2006.01) C07K 1/22 (2006.01) C07K 14/00 (2006.01) C07K 14/575 (2006.01) C07K 14/705 (2006.01) C07K 14/805 (2006.01) C07K 16/00 (2006.01) C07K 16/34 (2006.01) C12N 15/10 (2006.01) C12N 15/63 (2006.01) C12P 21/00 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2268397

The invention includes methods of isolating DNA and a protein encoded thereby, the protein being capable of binding to an antigen-bearing moiety. The methods comprise generating a DNA library expressing the protein in a virus vector. A magnetic label is added to cells expressing the antigen-bearing moiety and the cells are incubated with virus expressing the protein in the presence of an excess of unlabeled cells which do not express the antigen-bearing moiety to form a mixture, wherein the virus binds to the magnerically labeled cells. Cells having virus bound thereon are isolated from the mixture and DNA encoding the protein and the protein are obtained therefrom.

La présente invention concerne des procédés d'isolation d'ADN et d'une protéine par cet ADN codée, ladite protéine étant capable de liaison avec une fraction portant un antigène. Les procédés comportent la génération d'une bibliothèque d'ADN exprimant la protéine dans un vecteur viral. On ajoute un marquage magnétique aux cellules exprimant la fraction portant l'antigène et les cellules sont soumises à incubation avec le virus exprimant la protéine en présence d'un excès de cellules sans marquage n'exprimant pas la fraction portant un antigène, de façon à former un mélange dans lequel le virus se lie aux cellules marquées magnétiquement. Les cellules auxquelles est attaché un virus sont isolées du mélange, ce qui permet d'obtenir de l'ADN codant la protéine et ladite protéine.

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