Method for manufacturing high purified factor ó¨

Abstract

The present invention discloses a preparation method of a highly purified human blood coagulation factor IX. The human blood coagulation factor IX is prepared by performing anion exchange chromatography, cation exchange chromatography, heparin affinity chromatography to the material containing human blood coagulation factor IX (taken from human blood serum or recombined cell culture medium), which includes the period of passivating virus through S / D treatment (Solvent / Detergent treatment) or removing virus through nanofiltration. The highly purified safe coagulation factor IX preparation having a specific activity of above 150 IU / mg, with substantially undoped proteins can be prepared by the preparation method of the invention.

Description

A kind of preparation method of high-purity human coagulation factor IX technical field The present invention relates to a preparation method of high-purity human coagulation factor IX, in particular to a method by performing ion-exchange chromatography and affinity layer on a substance containing coagulation factor IX (taken from human plasma or recombined cell culture medium). In the stage of virus inactivation or virus removal by analysis, the safe high-purity coagulation factor IX with almost no adulterant protein and specific activity (specific activity) above 150IU / mg can be produced. Background technique Human coagulation factor IX is an indispensable glycoprotein in the blood coagulation process. If there is no coagulation factor IX or insufficient coagulation factor IX in the blood due to genetic factors or pathological factors, the coagulation process in the blood will be incomplete. , causing hemophilia B. Hemophilia B mainly occurs through heredity. One in 30...

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Problems solved by technology

And, there is also a more effective nanofiltration method that utilizes nanofiltration (nanofilter, 20nm), but when utilizing this nanofiltration, it is necessary to carry out a high-purity concentration process to the blood coagulation factor in advance, so as to prevent it from occurring in the use of non-high-purity When the prepared coagulation factor is subjected to nanofiltration, the filter is clogged and cannot be used

Although factor IX can be selectively isolated by monoclonal antibody (monoclonal antibody), since animal cells are required to produce antibodies, virus infection derived from animal cells cannot be ruled out

In addition, side effects caused by immunoglobulin (IgG) eluted with antibodies during the chromatography process cannot be ruled out

Also, the virus problem in isolating Factor IX from recombinant cell culture media cannot be completely overcome

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