PMID: 8987499Jan 1, 1995Paper

Stabile production of a thrombin resistant pro-urokinase derivative (PRO-UKS1) by Namalwa KJM-1 cells adapted to serum-free medium

Cytotechnology
S HosoiT Tamaoki

Abstract

Pro-UKS1 was designed as a thrombin-resistant derivative of pro-urokinase (pro-UK) by introducing a glycosylation site using site-directed mutagenesis. An expression plasmid for pro-UKS1, pMo1UKS1SEd1-5, was constructed and introduced into Namalwa KJM-1 cells (Hosoi et al., 1988), and cells resistant to G418 and Methotrexate (MTX) were obtained. Amongst them, the highest pro-UKS1 producer (resistant to 500 nM of MTX), clone 41-8, was selected and further characterized. Clone 41-8 was cultured in serum-free ITPSGF medium (Hosoi et al., 1988). Under the conventional conditions, the concentration of pro-UKS1 reached 26 micrograms ml-1. Addition of glucose and tri-iodothyronine (T3) improved productivity, and the maximal productivity of pro-UKS1 was 67 micrograms ml-1 day-1. In this conditioned medium, content of pro-UKS1 was above 80% of total proteins.

References

Nov 1, 1990·In Vitro Cellular & Developmental Biology : Journal of the Tissue Culture Association·M SatohS Sato
May 1, 1957·Biochimica Et Biophysica Acta·J PLOUG, N O KJELDGAARD

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