Biotechnological aspects of the production of human pro-kallikrein using the AcNPV-baculovirus-expression system.
We have investigated large scale production processes (up to 2 liters) of recombinant proteins using the baculovirus expression system in order to optimize the product yields. Experiments using cell lines of Spodoptera frugiperda (Sf9) and Mamestra brassicae (IZD-Mb0503) were performed to show the different production capacities of the cell lines. The influence of the infection at different cell densities is described. Beyond that, TC100-, IPL41- and serum-free IPL41-medium were compared to demonstrate their different capabilities of supporting cell growth and protein expression. Additionally, the inhibitory effect of FCS on the protease activity of kallikrein, which is produced in its zymogenic form, is discussed Improved production parameters are described, which enabled us to produce up to 8000 units of activated pro-kallikrein within 14 days using perfusion cultivation.
Fertig
G
G
Institute of Zoology, Technical University of Darmstadt, Germany.
Rahn
H P
HP
Angermann
A
A
Klöppinger
M
M
Miltenburger
H G
HG
D000818:Animals; D016367:Baculoviridae / Q000378:metabolism*; D001709:Biotechnology; D002417:Cattle; D002452:Cell Count; D002478:Cells, Cultured; D003470:Culture Media; D004792:Enzyme Precursors / Q000096:biosynthesis* / Q000378:metabolism; D006801:Humans; D007610:Kallikreins / Q000096:biosynthesis* / Q000378:metabolism; D009036:Moths / Q000166:cytology / Q000378:metabolism; D010477:Perfusion; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D014357:Trypsin
D003470:Culture Media; D004792:Enzyme Precursors; D011994:Recombinant Proteins; D007610:Kallikreins; C052910:prokallikrein, tissue; D014357:Trypsin