We are working on the ATH complex as a anticoagulant material. after production of the complex, we want to remove the unbounded AT and Heparin. for purification of this complex, the Butyle-agarose and DEAE sepharose were used in the reports. Is there another way other than this for remove unreacted AT and Heparin ? I will be appreciative for any help.
with best regards
Esfandyar
There are limitations because of the nature of your starting materials to a one-step chromatographic purification method.
You could try ammonium sulfate precipitation, this method requires you to determine the conditions empirically. You would expect that the complex would have a different precipitation point to the Heparin and/or antithrombin. the method is also relatively quick, cheap and easy.
However, if you are committed to a chromatographic methods, an alternative to DEAE usage would be able to use Size Exclusion chromatography to exclude unbound Heparin, but you would still need to use the Butyl-agarose (or any other Hydrophbic Interaction Chromatography media that you have ) to remove unbound antithrombin.
Of course you could try combining the SEC chromatography step with the ammonium sulfate step or any of the other published methods.
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