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Click to edit Master title,Click to edit Master text,Second level,Third level,Fourth level,Fifth level,*,血液制品层析工艺中,TOYOPEARL,填料,东曹,(,TOSOH,),公司,生命科学部门,Plasma Products Review,Application,on,Plasma Protein,Content,2,Protein,Concentration in Plasma,Indication,Albumin,40 g/L,Volume restoration after trauma,shock,burns,Alpha1 Proteinase Inhibitor,1.5 mg/mL,Hereditary emphysema,Anti-D IgG,Titer varies,a,Rh prophylaxis in pregnancy and childbirth,Antithrombin III,100 ug/mL,Antithrombin III deficiency,C1-inhibitor,170 ug/mL,Hereditary angioedema,Factor IX,10 ug/mL,Hemophilia B,Factor VIII,0.5 ug/mL,Factor VII deficiency,Factor XI,0.3 ug/mL,Hemophilia B,Factor XIII,30 ug/mL,Factor XIII deficiency,Fibrinogen,3 g/L,Tissue sealant component,Fibronectin,300 ug/mL,Wound healing,Hepatitis B IgG,Titer varies,a,Hepatitis immunity,Immunoglobulin G,Up to 12.5 g/L,Primary and secondary immune deficiency,Measles IgG,Titer varies,a,Measles protection and treatment,Protein C,4 ug/mL,Neonatal thrombosis,Rabies IgG,Titer varies,a,Rabies risk,Tetanus IgG,Titer varies,a,Tetanus protection and treatment,Thrombin,150 ug/mL,b,Tissue sealant component,Varicella Zoster IgG,Titer varies,a,Chicken pox protection,Von Willebrand Factor,10 ug,Von Willebrands disease,a,Titers(antibody concentrations)vary in ug/mL ranges.,b,As prothrombin,Plasma Proteins of Current Interest,3,Plasma Products in China,4,Plasma,precipitation,supernatant,Cryoprecipitate,Factor VIII,Fraction I,Centrifugation 4,o,C,Ethanol treatment,Centrifugation,supernatant,precipitation,Fibrinogen,Factor IX,Abtithrombin III,Cohn Cold Ethanol Fractionation Process,(Continued),5,precipitation,supernatant,Fraction IV,Fraction V,Ethanol treatment,Centrifugation,supernatant,precipitation,Haptoglobin,Abtithrombin III,Albumin,Fraction II+III,supernatant,precipitation,precipitation,Ethanol treatment,Centrifugation,Fraction II,Globulin,(for intramuscular),Globulin,(for intravenous),Ethanol treatment,Ethanol treatment,Centrifugation,Cohn Cold Ethanol Fractionation Process,6,Production Capacity of Plasma Proteins,Capacity;Million L/Y,Companies with yellow bar have,applied Toyopearl to their process.,7,CSL Bioplasma,Australia,Commonwealth Serum Laboratory(CSL),International pharmaceutical company based in Australia as CSL groups,All processes were applied,by chromatography for Cohn ethanol fractionation except Fraction 1 in 1994,The fourth largest supplier of plasma proteins(2002),Global company with manufacturing plant in US and Europe as well.,Consultant to Chinese plasma protein suppliers,8,Why CSL Bioplasma Applies Chromatography?,CSL Bioplasma describes chromatography as;,Highly cost effective process,High production efficiency(IVIG;5.5 g/L),High quality(Albumin;99%w/v),Achievable of automatic,closed-process,Easy extraction and separation of by-products (i.e.other plasma proteins),Advanced technique in bio-industry,9,Production Process for Plasma Proteins,Size Exclusion Chromatography,HW-55F,Ion-Exchange Chromatography,DEAE-650M,QAE-550C,SuperQ-650S,M,SP-650M,SP-550C,Hydrophobic Interaction Chromatography,Butyl-650M,C,Phenyl-650S,M,Affinity Chromatography,AF-Formyl 650M,AF-Heparin HC 650M,10,A,pplication of Plasma Products,11,Purification of Albumin,12,Purification of Albumin,by AIEC,Condition,Column:ToyoScreen SuperQ-650M(1mL),Eluent:(A)50 mM Tris-HCl,(B)50 mM Tris-HCl+0.5M NaCl,Gradient:(A)-(B)Linear 30min,Flow rate:1mL/min,Detection:UV(280nm),Sample:Human Albumin,Globulin 1g/L,Injection:100uL,13,Purification of Albumin,by CIEC,Condition,Column:ToyoScreen SP-650M(1mL),Eluent:(A)50 mM Acetic buffer(pH5.0),(B)50 mM Acetic buffer,+0.5M NaCl(pH5.0),Gradient:(A)-(B)Linear 30min,Flow rate:1mL/min,Sample:Human Albumin,Globulin 1g/L,Injection:100uL,14,Anion exchanger,Cation exchanger,Albumin,Globulin,Fraction V,Loading,Elution,Loading,Elution,Consideration,of Albumin Purification,15,Chromatography in plasma fractionation:benefit and future trends,T.Burnouf,J.Chromatgr.B 664(1995)3-15.,“TOYOPEARL DEAE-650M is used to produce a highly purified FVIII concentrate,.”,Purification of Factor VIII,16,Starting from Cohn fractionation,chromatography on TOYOPEARL,DEAE-650M results in a highly purified clotting factor for treatment of hemophilia A.,Method is licensed from Octapharma and includes two virus inactivation steps:solvent/detergent and heating.,Scale is hundreds of million units per year.,TOYOPEARL,DEAE-650M chromatography is the resin of choice.,Purification of Factor VIII,(Octanate),17,18,Condition,Column size:6.0mmID*4cmL(1.13ml),Mobile phase :Loading:0.4mg/ml AT-III,in 0.01M Tris-HCl,containing 0.15M NaCl pH7.5,Linear Velocity:300cm/min,Detection :UV(280nm),Resin AT-III binding capacity,(10%leakage,mg/ml-gel),New Heparin-650M3.9,Heparin Sepharose 6FF2.4,Heparin-650M1.9,Figure 1 Comparison of break through curve at 300cm/h,Comparison of dynamic,AT-III binding capacity at 300cm/h,Purification of AT-III,19,Condition,column size :6.0mmID*4cmL(1.13ml),Mobile phase :Loading:0.4mg/ml AT-III,in 0.01
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