The present invention describes methods and processes for the production
of proteins, particularly glycoproteins, by animal cell or mammalian cell
culture, illustratively, but not limited to, fed-batch cell cultures. The
methods comprise feeding the cells with D-galactose, preferably with feed
medium containing D-galactose, preferably daily, to sustain a sialylation
effective level of D-galactose in the culture for its duration, thus
increasing sialylation of the produced proteins. The methods can also
comprise at least two temperature shifts performed during the culturing
period, in which the temperature is lower at the end of the culturing
period than at the time of initial cell culture. The cell culture
processes of the invention involving two or more temperature shifts
sustain a high cell viability, and can allow for an extended protein
production phase. The methods can also comprise the delayed addition of
polyanionic compound at a time after innoculation. Supplementation of the
cultures with D-galactose, preferably in a feed medium, to sustain
galactose at sialylation effective levels in the cultures until the end
of a culture run reverses a decline in sialylation that accompanies
culture scale up, and is advantageous for large scale culturing
processes.