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KMID : 0380019940090010048
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Korean Journal of Biotechnology and Bioengineering
1994 Volume.9 No. 1 p.48 ~ p.54
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Kinetics of Converting Single Chain Urokinase Type Plasminogen Activator into Two Chain Plasminogen Activator in Cultivating HEK Cells with Low Serum Containing Medium
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Abstract
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A modified amidolytic assay and a fibrin plate method were used to accurately measure the concentration of single chain urokinase type plasminogen activator (scu-PA) and two-chain urokinase type plasminogen activator (tc-PA) in the spent media. 1.65 x 106( viable cells/ml) of maximum cell density and 1670 ( lU/ml) of scu-PA concentration were obtained in 1 % serum containing medium. he overall conversion ratio from scu PA to tc-PA was less than 10 %. In the results of batch cultivation in a spinner vessel, 4.43 x 10´( total cells/ml) of maximum cell density and 1560( lLUml) of scu PA concentration was observed. The maximum scu-PA concentration and specific scu-PA productivity were obtained in 1760( N/ml) and 3.13 x 10-´( lU/cell), respectively, from perfusion cultivation. The conversion ratios from batch, fed-batch and perfusion cultivations were less than 12%, which means that about 90 % of scu-PA secreted from the cells can be maintained during the cultivations.
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