Separation Reproducibility With the BioLogic Chromatography Systems ( + 1 M NaCl). All buffers were filtered...)

Separation Reproducibility With the BioLogic Chromatography Systems

+ 1 M NaCl). All buffers were filtered and degassed prior to use. For each run on the DuoFlow system, a 50 l sample loop was overfilled with 150 l of the protein sample using an Econo gradient pump. For the BioLogic LP system, 1 ml of sample was loaded though port C of the BioLogic LP buffer selection valve. The UV detector and conductivity monitor supplied with each system were used to monitor the UV signal and conductivity, respectively. Protocol details are summarized in Tables 1 and 2 and the system components used are listed in Table 3.

Results
The BioLogic DuoFlow and BioLogic LP systems were used to successfully purify a protein test mixture with excellent reproducibility. A total of 80 ion exchange runs were performed on the BioLogic DuoFlow QuadTec system as a queue of alternating anion exchange and equilibration runs (160 runs total). As shown in Figure 1, the 80 separations were virtually indistinguishable. The observed protein retention times (Table 4) demonstrate consistent sample introduction and pump performance. The standard deviation associated with the gradient slope, 4.88 0.01 mS/cm/min, indicates excellent pump and mixer performance, which is critical for discrete protein resolution. These results confirm that the BioLogic DuoFlow chromatography system provides high run-to-run reproducibility.

On the BioLogic LP, 25 consecutive separations were performed using its Multirun feature. As shown in Figure 2, the separations were highly reproducible and exhibited extremely consistent protein elution times (Table 4). Gradient performance was also highly reproducible with a
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( + 1 M NaCl). All buffers were filtered...)Separation Reproducibility With the BioLogic Chromatography Systems