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Performance Comparison of the Experion Automated Electrophoresis System and a Competing Automated System for Protein Analysis, Rev A

e was separated by SDSPAGE on 420% gradient gels (data not shown). On the other hand, the protein 200 plus assay detected an average of only 23 protein peaks from the same sample. Representative electropherograms are shown in Figure 2 to demonstrate the superior resolving capabilities of the Experion system.

The resolution of two symmetrical peaks can also be defined using a resolution value, Rs, which is defined by the equation Rs = 1.17(t2 t1) /(w1 + w2), where t1 and t2 = migration times of the protein peaks, and w1 and w2 = widths of the peaks at half height. Using this equation, Rs > 1.5 indicates that two adjacent peaks are baseline resolved.

To calculate the Rs that might theoretically be achieved with both systems, we used the migration time and peak width data collected from the Pro260 ladder separations as reference points. For this analysis, the Rs achievable by each system was calculated for each ladder peak (within each systems respective separation range) and a hypothetical protein 10% larger in MW but producing a peak of the same shape. Experion software exported the migration times and peak width data necessary for solving the equation; however, these data had to be manually determined for the bioanalyzer. Note that resolution numbers obtained were theoretical and that, in reality, the apparent resolution for chip separation is dependent on and limited by several factors. Such factors include the heterogeneity of a protein population, and modification of proteins in the sample, which can cause peak broadening and can alter protein migration.

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