The heparin ligand in the TSKgel Heparin-5PW affinity column can interact with sample molecules by cation exchange as well as affinity chromatography. Cation exchange interaction takes place through O-sulfate and N-sulfate groups. The elution strength is similar to TSKgel SP-5PW.

In vivo, heparin prevents blood clotting as it reacts with clotting agents. This tendency is taken advantage of by using heparin to separate blood clotting factors such as antithrombin III, Factor VII, Factor IX,etc., and anti-heparin proteins such as glycoproteins, endoglycosidase, hyaluronidase, lipases, growth factors, RNA polymerases, and other nucleic acid-binding proteins.