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Your protocol had problem.
I have ever purified a-glucosidase.
1. To Con A agarose affinity chromatography, the sugar in your protein binds to ConA, so you must use the buffer containing Methyl-a-glucose or Methyl-a-Mannose to
wash( for a-glucosidase).
2. The glycoprotein is easy to be solubilzed by high concentration of salt, thus you had better equilibrate the column with a buffer containing 40 mM hepes pH7.0, and
protease inhibitor, then use the same buffer containing 15 mM Methyl-a-Mannose to
wash.
