Methods and means for producing proteins with predetermined...

A - Human Necessities – 61 – K

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Details

A61K 38/18 (2006.01) C07K 14/075 (2006.01) C07K 14/505 (2006.01) C12N 15/00 (2006.01) C12N 15/65 (2006.01) C12N 15/85 (2006.01) C12N 15/86 (2006.01) G01N 33/68 (2006.01) C40B 40/10 (2006.01)

Patent

CA 2465007

The present invention provides methods for identifying, selecting and obtaining mammalian cells that are capable of producing proteinaceous molecules comprising predetermined post-translational modifications, wherein said post-translational modifications are brought about by the mammalian cell in which the proteinaceous molecule is expressed. Preferably, said predetermined post-translational modifications comprise glycosylation. The invention further provides methods for obtaining and producing proteinaceous molecules, using mammalian cells obtainable by a method of the present invention. Preferably said proteinaceous molecules comprise erythropoietin (EPO), since the effect of (recombinant) EPO depends heavily on the glycosylation pattern of the oligosaccharides present on the protein. Mammalian cells that have been obtained on the basis of their ability to produce proteins and/or post-translational modifications that are indicative for a predetermined post-translational modification that is desired are also provided. Preferably, said mammalian cells have neural characteristics and properties such that significant amounts of recombinant proteins can be produced that harbor 'neural- or brain-type' properties.

L'invention se rapporte à des procédés d'identification, de sélection et d'obtention de cellules mammifères capables de fabriquer des molécules protéiques présentant des modifications post-traductionnelles prédéterminées, lesdites modifications post-traductionnelles étant apportées par les cellules mammifères dans lesquelles s'exprime la molécule protéique. De préférence, lesdites modifications post-traductionnelles prédéterminées comprennent une étape de glycosylation. Cette invention porte aussi sur des procédés d'obtention et de fabrication de molécules protéiques au moyen de cellules mammifères pouvant être obtenues par un procédé selon l'invention. De préférence, lesdites molécules protéiques contiennent de l'érythropoïétine (EPO) puisque l'effet de l'EPO (recombinante) dépend en grande partie du schéma de glycosylation des oligosaccharides présents dans la protéine. L'invention en question concerne également des cellules mammifères qui ont été obtenues en fonction de leur capacité à fabriquer des protéines et/ou des modifications post-traductionnelles indiquant une modification post-traductionnelle prédéterminée voulue. De préférence, ces cellules mammifères possèdent des caractéristiques et des propriétés neuronales telles que des quantités significatives de protéines recombinantes peuvent être fabriquées afin de concentrer des propriétés <= neuronales ou de type cérébral >=.

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