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. Author manuscript; available in PMC: 2019 Jan 7.
Published in final edited form as: Biochemistry. 2017 Nov 28;56(49):6423–6433. doi: 10.1021/acs.biochem.7b00700

Figure 2.

Figure 2.

Two-step downstream purification scheme for rhTIMP-2– 6XHis. (A) Immobilized metal ion affinity chromatography (HisTrap column) for batchwise elution yields a single sharp elution peak using a step gradient of 20 mM (nonspecific) and 250 mM (specific, buffer B) elution. (B) Preparative reverse phase HPLC also yields a single Gaussian peak. The blue line indicates the % solvent B (acetonitrile, 0.1% TFA) composition of the mobile phase, with initial isocratic elution using 10% solvent B until 2 min and then a 10 to 60% solvent B gradient between 2 and 6 min. The column was then washed with a gradient of 60 to 100% solvent B between 6 and 10 min, before reequilibration with 10% solvent B.