Heparin-Sepharose affinity chromatography of human post-heparin plasma: Characterization of the Elution Pattern with Immunoelectrophoretic Methods

Abstract

Immunochemical methods including fused rocket-, crossed-, and tandem-crossed immunoelectrophoresis, have been used to characterize the elution pattern from heparin-Sepharose affinity chromatography of human post-heparin plasma. Hepatic triglyceride lipase (H-TGL) but not lipoprotein lipase (LPL) could be visualized with β-naphthyl acetate after immunoelectrophoresis. Two proteins were found to elute together with the lipolytic enzymes. The amino acid composition of fractions containing these proteins was nearly identical to that of anti-thrombin III. These results indicate that the removal of antithrombin III is the major problem in the purification of H-TGL and LPL from human post-heparin plasma by heparin-Sepharose affinity chromatography.

Key Words:

• antithrombin III
• enzyme antibodies
• lipoprotein lipase
• hepatic triglyceride lipase