Methods of Plasma Fractionation: Solid-Phase Reagents

摘要

The solid-phase ethylene/maleic anhydride PEs E-5 and E-100 were used to develop a flexible integrated system of batch methods for the fractionation of albumin and gamma globulin as well as the clinically important coagulation factors. Each method for the isolation and purification of the fractions or a modification of it could also be used independently or could be added onto another method or methods. Each fraction was prepared in three steps: adsorption on solid-phase PEs, elution from the PEs, and concentration by ultrafiltration or by use of PEG or both. Since the methods required no organic solvent but relied primarily on electrostatic bonding, they were essentially nondenaturing. Solid-liquid phase separation was easily accomplished by centrifugation or filtration. The yield and purity of each fraction was as follows: (1) coagulation factors II, VII, IX and X concentrates 63% and 200-fold purified; (2) AHF 29-50%, depending on the method, and 52-fold; (3) vWF 43% and 71-fold; (4) PPF, as albumin, 93% yield and 92% pure; (5) albumin 92% yield and 98.5% pure; and (6) gamma globulin 92% yield and 97.5% pure. During the preparation of gamma globulin the E-100 also tended to adsorb and remove the HBsAg and infectivity, when present. Toxicity studies in animals using aqueous extracts of PE E-5 and PE E-100 and with purified AHF, albumin and gamma globulin showed no abnormal results.