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. Author manuscript; available in PMC: 2008 Sep 1.
Published in final edited form as: Protein Expr Purif. 2007 Apr 25;55(1):53–68. doi: 10.1016/j.pep.2007.04.013

Figure 10.

Figure 10

Results from the automated two-step purification of MHT238Δ. A, absorbance and gradient profiles for the two-step purification. The lower solid line shows the absorbance profile during the 2nd cycle of sample injection, wash, and elution steps and the 3rd cycle of sample injection for the IMAC purification. The upper solid line shows the absorbance profile for the 1st and 2nd cation exchange steps. The dashed lines indicate the percentage mixture of buffer B during the IMAC step and buffer D during the cation exchange step. The area of elution peaks cannot be directly compared due to the different flow rates during IMAC and cation exchange elution steps. B, characterization of the purification results by SDS-PAGE. Lane 1 is the cell-free lysate and lanes 2 and 3 are the IMAC and cation exchange waste products. The waste products were concentrated using a 10-kDa molecular weight cut off centrifugal concentrator to the same volume as the cell lysate for easier visual comparison. Lanes 4 through 9 are the purified TEV protease obtained from purification cycles 1 through 6. Lane 10 shows a higher loading (50 μg) of the sample from lane 5 (10 μg) and lane 11 contains 312 ng of bovine serum albumin standard run on the same gel as lane 10.