| 摘要 |
PROBLEM TO BE SOLVED: To develop an FVII composition with an extremely reduced immunogenicity and with increased viral safety.SOLUTION: A process of extraction and purification of transgenic FVII contained in milk of a transgenic animal comprises: a step (a) of extracting the FVII from the milk, the anion of a soluble salt being selected for its capability to release the Factor VII, the Factor VII being then present in a liquid phase; a step (b) of separating a protein-enriched liquid phase from the precipitate of calcium compounds, the liquid phase being further separated in a lipidic phase and an aqueous nonlipidic phase containing the protein; a step (c) of subjecting the aqueous nonlipidic phase to an affinity chromatography step, using an elution buffer based on a phosphate salt in a predetermined concentration; and a step (d) of subjecting the eluate of Factor VII, obtained according to the step (c), to two or three chromatography steps on weak basic type anion exchange columns using buffers suitable for successive elutions of the Factor VII retained on the columns. |
