Affinity fractionation of heparin on immobilized Concanavalin A

摘要

A highly discriminating technique utilizing Concanavalin A, immobilized on a solid substrate, e.g. sepharose 4B, for the fractionation of heparin is disclosed. The heparin to be fractionated is incubated with antithrombin whereby a fraction complete with the antithrombin and the whole is then either passed through a column of the immobilized Concanavalin A or slurried with same. In both modes that fraction of the heparin complexed with the antithrombin is selectively absorbed by the immobilized Concanavalin A and constitutes the anticoagulant active fraction. That heparin which complexes with the antithrombin can be further fractionated to yield yet more highly active fractions by complexing the heparin in stages, adding an increment of a molar amount of antithrombin in each stage. A high molecular weight fraction (18,000-22,000 daltons) or a low molecule weight fraction (6,000- 8,000 daltons), isolated from the heparin of animal tissue origin by chromatography, are the preferred heparin preparations for complexing with the antithrombin.