High-performance affinity chromatography heparin

A. S. Clairbois, D. Letourneur, D. Muller, and J. Joze-fonvicz. High-performance affinity chromatography for the purification of heparin-binding proteins from detergent-solubilized smooth muscle cell membranes, J. Chromatogr. B 706 55 (1998). 

In the present paper, we report high-performance affinity chromatography of thrombin in presence of AT III and Hep, using two types of resins as stationary phases either heparin-like PSSO or AT Ill-like PAOM. In order to differentiate their mechanisms of interaction with thrombin, we examined the chromatographic behavior of thrombin in the presence, or in the absence of AT III and/or heparin. Finally, thrombin was injected on the columns at low ionic strength. The desorption of bound thrombin from the two solid surfaces was then carried out using AT III, heparin and the AT Ill-Hep complex, to elucidate the specificity of the interactions involved. 

Heparinlike sulfated dextran derivatives, like carboxymethyldextran benzylamide sulfonates (CMDBS), have been immobilized on silica beads. By high-performance liquid affinity chromatography (HPLAC), they allow a good recovery of thrombin, with a yield of 80%. The affinity constant was estimated K lO M ) and was found superior to the value obtained between thrombin and heparin. On the contrary, the affinity of dextran derivatives for ATIII is estimated at a lower value than that of heparin [17],... 

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