Process for cell culturing by continuous perfusion

摘要

The invention relates to a process for the culturing of cells by continuous perfusion culturing of a cell culture comprising cell culture medium and cells, wherein cell culture medium is added to the cell culture, the cell culture is circulated over a filter module comprising hollow fibers resulting in an outflow of liquid having a lower cell density than the cell culture and the flow within the filter module is an alternating tangential flow. Preferably, culture medium is added at a particular perfusion rate and/or biomass is removed form the culture at least once. The method is especially suitable for the culturing of aggregating cells. The invention also relates to such a process wherein a biological substance, preferably an antibody, is produced by the cells, which biological substance may be further purified in downstream processing.

主权项

1. A method for limiting cell aggregation during cell culture to a level wherein no more than 5% of the animal cells in suspension in the culture comprise aggregates of at least 5 cells, comprising:
culturing of a cell culture comprising cell culture medium and animal cells that easily or inherently form aggregates during culturing, wherein the culturing comprises: (a) circulating the cell culture through a filter module comprising hollow fibers, in an alternating tangential flow resulting in an outflow of cell culture liquid through the pores of the filter module having a lower animal cell density per ml than the cell culture prior to circulating through the filter module; and (b) adding cell culture medium to the cell culture, and wherein no more than 5% of the animal cells in the culture form aggregates in suspension of at least 5 cells during the culturing, and wherein the culturing is continued until animal cells are present in the cell culture at a density of at least 80×106 viable animal cells/ml.