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Expression of human factor X with normal biological activity in human embryonic kidney cells

โœ Scribed by Deog Joong Kim; Harold L. James

Publisher
Springer Netherlands
Year
1994
Tongue
English
Weight
396 KB
Volume
16
Category
Article
ISSN
0141-5492

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โœฆ Synopsis

Human embryonic kidney cells (293) were transfected with a construct containing human factor X cDNA and selected for G418 resistance. The level of expression of recombinant factor X in serum-free medium was 4 to 5 pg/ml. Purified recombinant factor X had a molecular size identical to that of normal plasma factor X. Amino-terminal sequencing revealed normal processing cleavages. The rcarboxy Glu and 13-OH Asp content of the recombinant factor X was close to 90% of the expected levels of these post-translational residues. The specific activity of recombinant factor X was about 95% of that of plasma factor X in three plasma-based clotting assays. This report demonstrates that 293 cells can produce a high level of biologically active factor X and describes a visual criterion for verifying the transfection process.

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