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Although a limitation is that halogenated compounds containing chloride ions (e.g., Tris-HCl, NaCl) cannot be used in the lysis or binding buffers, the system offers the advantage of one-step purification (~1 h) with high specificity, as has been reported in several applications involving recombinant protein expression and purification using this system [43,44,165,166]

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Unbound protein is washed through the column with five column volumes of Buffer N1