Patent ID: 6103500
Filing Date: 2000-08-15
Classification: A61K,C07K,C12N

Abstract:
A method for producing a factor VIIa/TF/Xa binding protein, said method comprising:(a) transforming yeast cells with a vehicle, said vehicle comprising a first nucleotide sequence encoding the factor VIIa/TF/Xa binding protein, said factor VIIa/TF/Xa binding protein selected from the group consisting of TFPI and a TFPI mutein, wherein the N-terminal amino acid sequence of the factor VIIa/RF/Xa binding protein is SEQ ID NO: 7, said first nucleotide sequence being immediately preceded in frame by a second nucleotide sequence encoding ubiquitin, the first and second nucleotide sequences together encoding a fusion peptide;(b) incubating the transformed yeast cells under conditions favorable for production of the factor VIIa/TF/Xa binding protein, wherein the factor VIIa/TF/Xa binding protein is retained within the yeast cell;(c) preparing an insoluble fraction of the transformed yeast cells containing the factor VIIa/TF/Xa binding protein; and(d) recovering the factor VIIa/TF/Xa binding protein from the insoluble fraction.