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The column was washed with 6 bed volumes of wash buffer (50 mM sodium phosphate, 300 mM NaCl and 100 mM imidazole, pH 7.4) and the target protein was eluted with 6 bed volumes of elution buffer (50 mM sodium phosphate, 300 mM NaCl and 300 mM imidazole, pH 7.4)