I currently have access to vectors that will express either ScFv anti-His, or ScFv anti- His fused to Alk. Phos. As we can express and purify unlimited amounts of these anti-his antibody fragments, I would like to use them in an immunoaffinity column in order to purify a his tagged polypeptide. Has anyone used ScFv fragments for immunoaffinity chromatography? if so what would be the best method of immoblilising the ScFv to a solid phase substrate so that the epitope binding region is free for interactions with the antigen? Help much appreciated
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