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

analysis reveals that this assay has a sensitivity comparable to that of colloidal Coomassie Blue G-250 stain, and enables the detection of impurities that are present in a sample at levels as low as 0.10.2% of the target protein concentration.

Since the protein 200 plus assay could not detect the 2.5 ng/l CA sample, an electropherogram showing an overlay of the 2.5, 10, and 50 ng/l CA samples was used to estimate the relative sensitivity of both assays (Figure 5). The Experion Pro260 analysis of the 2.5 ng/l sample generated relative fluorescence units similar to those obtained for the 10 ng/l sample analyzed with the protein 200 plus assay. The same trend was observed for the 10 ng/l and 50 ng/l samples. These data imply an approximately 4-fold greater sensitivity of the Pro260 assay over the protein 200 plus assay, though working near the limit of detection of any instrument or assay is rarely advisable.

In the above analyses, bovine CA was used in determining the sensitivity of both systems. However, when other protein samples are used, various factors may influence the overall assay sensitivity for either system. First, sensitivity may vary depending on the staining efficiency of a particular protein, as discussed previously. Thus, a protein with poor affinity for the stain would have to be present at concentrations above 2.5 ng/l to be detected above the baseline. Second, the shape of a peak may affect the lower detection limit, as broader peaks are more difficult to identify above the baseline. Finally, and probably most significantly, the buffer composition plays a large role in setting the limit of detection for a protein sample. Samples are loaded into the separation channel of
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