Affinity Chromatography on Immobilised Adenosine 5′‐monophosphate

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Abstract
The binding of various enzymes to two chemically defined affinity adsorbents, P1‐(6‐aminohexyl)‐P2‐(5′‐adenosine)‐pyrophosphate‐Sepharose and N6‐(6‐aminohexyl)‐5′‐AMP‐Sepharose, has been compared. The results have been interpreted in terms of the enzyme‐nucleotide interaction. The effect of ligand concentration on the nature and strength of enzyme binding to immobilised 5′‐AMP has been investigated and shown to be a critical paramcter. Dilution of the immobilised ligand with Sepharose or preparation of low ligand concentration adsorbents, can be utilised to facilitate elution of tightly bound enzymes. Conversely, significant increases in the binding and capacity of affinity adsorbents can be achieved by increasing the ligand concentration. The binding of lactate dehydrogenase in response to increasing ligand concentrations was not a simple function. The data have been interpreted in the light of current concepts on affinity chromatography.