Irx-2 modified manufacturing process

Details Irx-2 modified manufacturing process Irx-2 modified manufacturing process

C - Chemistry, Metallurgy

12

P

C12P 21/02 (2006.01) C12N 5/078 (2010.01) C07K 14/52 (2006.01) C12M 1/00 (2006.01) C12M 3/00 (2006.01) C12N 1/00 (2006.01) C12N 15/10 (2006.01) C12P 1/00 (2006.01) C12N 13/00 (2006.01)

Patent

CA 2758834

A highly efficient method of making a primary cell derived biologic by purifying mononuclear cells (MNCs) in a automated cell processor to remove contaminating cells by loading leukocytes onto lymphocyte separation medium (LSM) and centrifuging the medium to obtain purified MNCs, storing the MNCs overnight in a closed sterile bag system, stimulating an induction mixture of the MNCs with phytohemagglutinin (PHA) or other mitogen and ciprofloxacin in a scalable cell culture device and producing a primary cell derived biologic from the MNCs, removing the mitogen from the induction mixture by filtering, incubating the induction mixture, clarifying the induction mixture by filtering to obtain a primary cell derived biologic supernatant, and clearing the primary cell derived biologic supernatant from adventitious agents by anion exchange chromatography, filtration. A closed system prevents contamination of the resulting primary cell derived biologic. An automated method of purifying cells. A method of scalably inducing cells.

Linvention concerne un procédé haute efficacité de fabrication dun agent biologique issu de cellules primaires par purification de cellules mononucléaires (CMN) dans un processeur de cellules automatisé, de sorte à éliminer les cellules contaminantes. Ce procédé consiste : à charger des leucocytes sur un support de séparation de lymphocyte et à centrifuger le support pour obtenir des cellules mononucléaires purifiées; à stocker les CMN pendant une nuit dans un système de sac stérile fermé; à stimuler un mélange dinduction des CMN avec de la phytohémagglutinine (PHA) ou un autre mitogène et de la ciprofloxacine dans un dispositif de culture cellulaire échelonnable et à produire un agent biologique issu de cellules primaires à partir des CMN; à éliminer le mitogène du mélange dinduction par filtration; à incuber le mélange dinduction; à clarifier ledit mélange par filtration pour obtenir un surnageant dagent biologique issu de cellules primaires; et à débarrasser ledit surnageant des agents adventifs par chromatographie par échange danions et filtration. Un système fermé empêche la contamination de lagent biologique résultant. Linvention concerne également un procédé automatisé de purification de cellules et un procédé dinduction échelonnable de cellules.

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