Method for fractionating plasma proteins using peg and ion-exchangers

Claim 1

A METHOD OF FRACTIONATION PLASMA PROTEINS, COMPRISING THE FOLLOWING STEPS: A. REMOVING BLOOD CORPUSCLES AND CELL FRAGMENTS FROM THE PLASMA IN THE BLOOD; B. PRECIPITATING THE GLOBULINS IN THE PLASMA WITH POLYETHYLENE GLYCOL HAVING A MOLECULAR WEIGHT OF APPROXIMATELY 6000 IN A CONCENTRATION OF 10 TO 15% BY WEIGHT AND AT A PH OF 6.5 TO 8.0 AT ROOM TEMPERATURE; C. CENTRIFUGING OUT ALL OF THE PRECIPITATE FROM THE REMAINING SOLUTION; D. DISSOLVING THE PRECIPITATE AT PH 5.8 IN 0.05 M SODIUM ACETATE-ACETIC ACID BUFFER AND CENTRIFUGING OUT UNDISSOLVED FIBRIOGEN, PLASMINOGEN AND IGM; E. ADSORBING THE GLOBULINS FROM THE SOLUTION CONTAINING B-LIPOPROTEINS, A-MACROGLOBULINS, LIPIDS, PRECITITATED ALBUMIN AND IGG OBTAINED IN STEP (D) ON CARBOXY METHYL DEXTRAN CATION EXCHANGER AT A PH OF 5.8, THEREBY LEAVING IN THE SOLUTION THE B-LIPOPROTEINS, A-MACROGLOVULINS, LIPIDS, PRECIPITATED ALBUMIN AND A LESSER AMOUNT OF IGG; AND ELUTING THE CATION EXCHANGER WITH 0.2 M SODIUM CHLORIDE IN 0.05 M PHOSPHATE BUFFER AT PH 7.5 TO 8.0 TO 6.6; F. PRECIPITATING THE ELUTE FROM STEP (E) WITH ADDITIONAL POLYETHYLENE GLYCOL AT PH 6.5 TO 8.0, CENTRIFUGING AND DISSOLVING THE PRECIPITATE IN 0.2 M PHOSPHATE BUFFER AT PH 6.6; G. ADSORBING ALL THE GLOBULINS IN THE SOLUTION FROM STEP (F) EXCLUDING IGG ON AN ANION EXHCANGER AT PH 6.6; H. COOLING THE IGG SOLUTION TO -0.3*C. AND PRECIPITATING IGG WITH ETHANOL IN A CONCENTRATION OF 25% BY VOLUME OF THE TOTAL SYSTEM AT -5* TO -10*C, SAID USE OF ETHANOL BEING THE FIRST USE OF ETHANOL IN SAID PROCESS; I. SEPARATING THE PRECIPITATE, SUSPENDING AND WASHING THE 25% BY VOLUME ETHANOL AT PH 7.0 AT -5* TO -10*C., AND THEN RENEWING CENTRIFUGING; J. DISSOLVING IN 0.1 M GLYCINE AND STERILE-FILTERING; AND K. FREEZE-DRYING AND PREPARING AT 16 TO 16.5% IGG SOLUTION IN 0.3 M GLYCINE FOR INJECTION.

Claim 2

A METHOD OF FRACTIONATION PLASMA PROTEINS, COMPRISING THE FOLLOWING STEPS: A. REMOVING BLOOD CORPUSCLES AND CELL FRAGMENTS FROM THE PLASMA IN THE BLOOD; B. PRECIPITATING THE GLOBULINS IN THE PLASMA WITH POLYETHYLENE GLYCOL HAVING A MOLECULAR WEIGHT OF APPROXIMATELY 6000 IN A CONCENTRATION OF 10 TO 15% BY WEIGHT AND AT A PH OF 6.5 TO 8.0 AT ROOM TEMPERATURE; C. CENTRIFUGING OUT ALL OF THE PRECIPITATE FROM THE REMAINING SOLUTION; D. DISSOLVING THE PRECIPITATE AT PH 5.8 IN 0.05 M SODIUM ACETATE-ACETIC ACID BUFFER AND CENTRIFUGING OUT UNDISSOLVED FIBRIOGEN, PLASMINOGEN AND IGM; E. ADSORBING THE GLOBULINS FROM THE SOLUTION CONTAINING B-LIPOPROTEINS, A-MACROGLOBULINS, LIPIDS, PRECITITATED ALBUMIN AND IGG OBTAINED IN STEP (D) ON CARBOXY METHYL DEXTRAN CATION EXCHANGER AT A PH OF 5.8, THEREBY LEAVING IN THE SOLUTION THE B-LIPOPROTEINS, A-MACROGLOVULINS, LIPIDS, PRECIPITATED ALBUMIN AND A LESSER AMOUNT OF IGG; AND ELUTING THE CATION EXCHANGER WITH 0.2 M SODIUM CHLORIDE IN 0.05 M PHOSPHATE BUFFER AT PH 7.5 TO 8.0 TO 6.6; F. PRECIPITATING THE ELUTE FROM STEP (E) WITH ADDITIONAL POLYETHYLENE GLYCOL AT PH 6.5 TO 8.0, CENTRIFUGING AND DISSOLVING THE PRECIPITATE IN 0.2 M PHOSPHATE BUFFER AT PH 6.6; G. ADSORBING ALL THE GLOBULINS IN THE SOLUTION FROM STEP (F) EXCLUDING IGG ON AN ANION EXHCANGER AT PH 6.6; H. COOLING THE IGG SOLUTION TO -0.3*C. AND PRECIPITATING IGG WITH ETHANOL IN A CONCENTRATION OF 25% BY VOLUME OF THE TOTAL SYSTEM AT -5* TO -10*C, SAID USE OF ETHANOL BEING THE FIRST USE OF ETHANOL IN SAID PROCESS; I. SEPARATING THE PRECIPITATE, SUSPENDING AND WASHING THE 25% BY VOLUME ETHANOL AT PH 7.0 AT -5* TO -10*C., AND THEN RENEWING CENTRIFUGING; J. DISSOLVING IN 0.1 M GLYCINE AND STERILE-FILTERING; AND K. FREEZE-DRYING AND PREPARING AT 16 TO 16.5% IGG SOLUTION IN 0.3 M GLYCINE FOR INJECTION.