Extraction of integral membrane proteins

Details Extraction of integral membrane proteins Extraction of integral membrane proteins

C - Chemistry, Metallurgy

07

K

C07K 14/285 (2006.01) C07K 1/34 (2006.01)

Patent

CA 2370694

A process is described for extracting gram-negative integral membrane proteins from bacteria or bacterial host cells containing a recombinant vector by differential detergent tangential flow diafiltration. This process has several advantages over alternate processes. First, it combines the clarification and extraction processes into one unit operation. The product is extracted from the cells and it is separated from cell debris with only one continuous diafiltration process. Second, the membrane proteins are extracted in a semi-purified state, which simplifies the downstream processing steps. Third, this process is very scalable because the only requirement is that the surface area of the membranes be increased proportionally with the amount of cells.

L'invention concerne un procédé d'extraction de protéines membranaires entières Gram négatif, à partir de bactéries ou cellules hôtes bactériennes contenant un vecteur de recombinaison, ce procédé consistant en une diafiltration différenciée à écoulement tangentiel, au moyen d'un détergent. Ce procédé comporte plusieurs avantages sur des autres procédés. D'abord, il combine les processus de clarification et d'extraction en une seule opération, le produit étant extrait des cellules et séparé des débris de celles-ci au moyen d'un seul processus de diafiltration en continu. Ensuite, les protéines membranaires sont extraites à l'état semi-purifié, ce qui simplifie les étapes de traitement en aval. Troisièmement, ce procédé est très facilement modifiable, étant donné que la seule règle à respecter est l'augmentation de la surface des membranes proportionnellement à la quantité de cellules.

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