Extraction of heterologous insoluble proteins from bacteria

Details Extraction of heterologous insoluble proteins from bacteria Extraction of heterologous insoluble proteins from bacteria

C - Chemistry, Metallurgy

12

P

C12P 21/00 (2006.01) C07K 1/14 (2006.01) C07K 14/54 (2006.01) C12N 15/70 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2192184

An insoluble mammalian protein is ex- tracted from transformed bacteria express- ing the mammalian protein while avoiding irreversible insolubilization of bacterial host proteins by homogenizing the fermentation broth, centrifuging the homogenized broth and removing the supernatant liquid from the inclusion body containing pellet. In another embodiment, the pH of the homogenized broth is adjusted to 2.0 prior to centrifuga- tion. The acidified broth is then centrifuged, and the pellet is resuspended in buffer, ho- mogenized again, and the inclusion body is isolated by centrifugation.

Une protéine mammalienne insoluble est extraite de bactéries transformées exprimant la protéine mammalienne, tout en évitant l'insolubilisation irréversible des protéines hôtes bactériennes par homogénéisation du bouillon de fermentation, centrifugation du bouillon homogénéisé et élimination du liquide surnageant dans le corps d'inclusion contenant le culot de centrifugation. Dans une autre réalisation, le pH du bouillon homogénéisé est amené à 2,0 avant centrifugation. Le bouillon acidifié est alors centrifugé, puis le culot est remis en suspension dans une solution tampon, homogénéisé à nouveau, et le corps d'inclusion est isolé par centrifugation.

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