Problems with serum-free production of antithrombin III regarding proteolytic activity and product quality.
Human antithrombin III (AT-III) was produced using a recombinant BHK-21 cell line with a microcarrier culture in spinner flasks. Cells were cultivated for the first 4 days in a medium containing 10% fetal calf serum (FCS). Afterwards, the medium was exchanged and production of AT-III occurred at high cell numbers in a serum-free medium. The product was determined by an immunoassay and further analysed after isolation from the culture medium. During cultivation, high proteolytic activity was detected which caused a considerable product decomposition. Furthermore, a higher level of non-glycosylated AT-III was found after serum-free production.
Teige
M
M
Institut für Pflanzengenetik und Kulturpflazenforschung, Gatersleben, Germany.
Weidemann
R
R
Kretzmer
G
G
D000818:Animals; D000990:Antithrombin III / Q000096:biosynthesis* / Q000592:standards; D002460:Cell Line; D006224:Cricetinae; D016895:Culture Media, Serum-Free; D010450:Endopeptidases / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis*
D016895:Culture Media, Serum-Free; D011994:Recombinant Proteins; D000990:Antithrombin III; D007770:L-Lactate Dehydrogenase; D010450:Endopeptidases