The invention belongs to the technical field of affinity chromatography, and provides an efficient and simple protection method of an ADP aglucon chromatography media. The method comprises the following steps: adding a chelating agent into flavoprotein TrxR crude enzyme, so as to chelate Mg<2+> in the crude enzyme, inhibit DNase activity and protect ADP aglucon; balancing a 2'5'ADP Sepharose affinity column through a TE buffer solution, adding the TrxR crude enzyme containing the chelating agent intothe column to ensure that TrxR is combined with the ADP aglucon, washing miscellaneous proteinnonspecifically combined on the column with the TE buffer solution, eluting the TrxR combined with the ADP aglucon with TE+0.5M NaCl, treating the column with a regulation buffer solution to perform regeneration on the column, and then immediately balancing the column subjected to regeneration with the TE buffer solution. The method has the advantages that an operation method is simple, the chelating agent is cheap and east to get, the economic cost is low, the protection effect on the ADP aglucon is obvious, and the practicability is high.