Large scale methods of producing adenovirus and adenovirus...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 7/00 (2006.01) A61K 31/395 (2006.01) C12N 7/01 (2006.01) C12N 7/02 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2478932

A process for large scale virus production is disclosed, especially large scale adenovirus production. The methods described are preferably adapted to suspension culture of mammalian host cells in a large scale bioreactor where gas sparging becomes essential to provide adequate aeration through the duration of the culture. This methodology includes an elevated concentration of a compound which protects host cells from the shearing effects of gas sparging and agitation as well as utilizing virus seed stocks which have been generated free of cell lysis reagents. The invention also relates to methods of producing these virus seed stocks, which are scaleable and the resultant unclarified virus seed stocks which are concentrated to reduce storage volume for infection of large scale culture and which are free of cell lysis components, such as the detergent Triton X-100 or Polysorbate 80.

La présente invention concerne un processus de production à grande échelle de virus et plus spécifiquement la production à grande échelle d'adénovirus. Les procédés décrits sont de préférence adaptés à la culture en suspension de cellules hôtes de mammifères dans un bioréacteur à grande échelle dans lequel le barbotage du gaz devient essentiel pour assurer une aération adéquate pendant toute la durée de la culture. Cette méthodologie comprend l'utilisation d'une concentration élevée d'un composé qui protège les cellules hôtes des effets de cisaillement du barbotage du gaz et de l'agitation ainsi que l'utilisation de collections de semence de virus qui ont été produites sans réactifs de lyse cellulaire. Cette invention concerne également des procédés de production de ces collections de semence de virus, qui sont échelonnables et les collections de semence de virus non clarifiés résultantes qui sont concentrées pour réduire le volume de stockage pour infecter une culture à grande échelle et qui sont dépourvues de constituants de lyse cellulaire, tels que le détergent Triton X-100 ou Polysorbate 80.

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