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the column was washed with 20 ml equilibration solution, and elution was performed with a linear pH gradient from 3.2 to 5.0 (0.85 % (w/w) tri-sodium citrate, 0.33 % (w/w) succinic acid, 0.6 % (w/w) di-sodium hydrogen phosphate di-hydrate, 45 % (w/w) ethanol) followed by 60 ml of isocratic elution at pH 8.0 (0.85 % (w/w) tri-sodium citrate, 0.33 % (w/w) succinic acid, 0.6 % (w/w) di-sodium hydrogen phosphate di-hydrate, 45 % (w/w) ethanol, pH 8.0)