Factors Concerning the Effectiveness of Affinity Chromatography Using Immobilized Cofactors

Abstract

Considerable interest has been fostered in the development and application of immobilized “general ligands” such as the adenine-containing coenzymes. This interest derives from the usefulness of such derivatives in affinity chromatography (1–14) and from their potential applications in enzyme reactors (5,6). The chemical complexity of the adenine nucleotide coenzymes has until recently hampered the development of methods suitable for their immobilization. Nevertheless, despite acute synthetic problems, several chemically characterized polymers have been described (7–9). Most methods have concentrated on derivatization of the adenine moiety of the coenzyme and in particular in substitution at the exocyclic N⁶-amino group. X-ray data from several dehydrogenases has shown that this N⁶-amino group protrudes out of a hydrophobic pocket in which the adenosine moiety of the coenzyme is bound (10).