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- W2365987341 abstract "Objective To control the quality of pilot process for purification of human coagulation factor Ⅷ(FⅧ) by PEG precipitation combined with ion exchange chromatography.Methods The test samples of FⅧ after pilot purification were determined for coagulation activity by coagulation assay,and for total protein content by BCA method,based on which the specific activity was calculated.The albumin(ALB),IgG,fibrinogen(FIB)and fibronection(FNC)contents were determined by turbidimetry,while the FⅧ and von Willebrand factor(vWF) antigen contents by double antibody sandwich ELISA,and the specificity of F Ⅷ by SDS-PAGE and Western blot.The residual PEG,Tween 80 and tr(inbutyl)phosphate(TNBP)contents were determined according to the requirements in Chinese Pharmacopeia(Volume Ⅲ,2010 edition).Results Most of impurities were removed by chromatography.The specific activity of F Ⅷ after purification increased by about 20 folds as compared with that before purification,which reached 66.92 IU / mg.ALB,IgG,FIB and FNC were basically removed by PEG precipitation and DEAE chromatography,with few losses of FⅧ and activity.The main components in bulk were FⅧ and vWF.A specific band with relative molecular mass between 95 000 and 72 000 was observed on the elution peak of DEAE chromatography,which was consistent with those of FⅧ light chain(Mr 80 000)and freeze-dried national standard for FⅧ.Both the residual contents of PEG,Tween 80 and TNBP met the requirements in Chinese Pharmacopeia(Volume Ⅲ,2010 edition).Conclusion The FⅧ with high purity and high specificity may be prepared by the pilot purification process." @default.
- W2365987341 created "2016-06-24" @default.
- W2365987341 creator A5054803337 @default.
- W2365987341 date "2013-01-01" @default.
- W2365987341 modified "2023-09-28" @default.
- W2365987341 title "Quality control of pilot purification process of human coagulation factor VIII" @default.
- W2365987341 hasPublicationYear "2013" @default.
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