| abstract |
A chromatographic method and apparatus for purifying proteins generally, and monoclonal antibodies from ascites fluids specifically, is disclosed. The method comprises contacting a solution containing proteins to be purified with a porous, substantially nonswellable sheet which has hydrated crystalline hydroxylapatite particles physically immobilized therein, in the presence of a pH buffer, and then eluting purified protein fractions, such that the flow rate and back pressure do not change significantly throughout the method. One preferred apparatus comprises inlet and outlet seals (10, 15), spiral- wound or core-wrapped chromatographic separation medium (60) comprising hydrated crystalline hydroxylapatite particles physically immobilized in a porous, substantially nonswellable sheet, and a Porous central support (40) upon which the sheet is wrapped. Another apparatus comprises single or stacked chromatographic separation medium comprising hydrated crystalline hydroxylapatite particles physically immobilized in a porous, substantially nonswellable sheet. <IMAGE> |