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

the chip by an electrokinetic injection process; therefore, the amount of protein injected depends on the ionic strength of the buffer in which the sample is solubilized. A buffer conductivity similar to that of 1x PBS is recommended. If the protein sample has a higher conductivity, then less protein will be injected and the limit of detection will rise. The opposite should also be true for samples of low conductivity, the limit of detection will likely decrease, thereby enhancing sensitivity.

Effects of Salt
The fact that the sensitivity of an assay decreases with increasing sample conductivity may be problematic because many chromatographic procedures involve eluting fractions with a salt gradient, or in a stepwise fashion with increasing salt concentration. It is, therefore, important that the protein assay used to analyze these chromatographic fractions be able to handle varying levels of salt without affecting the accuracy and reproducibility of sizing or quantitation.

To examine the effect of salt concentration on the ability of each system to correctly determine the MW and concentration of each CA sample, alternating wells of each chip were loaded with 100 ng/l CA in 10 mM sodium phosphate buffers (pH 7.4) containing either 0.15 M NaCl (1x PBS), 0.25 M NaCl, or 0.5 M NaCl. With both systems, no obvious salt-related effects on the accuracy and reproducibility of sizing were observed (Table 3). Increasing salt concentrations did, however, have a pronounced effect on the accuracy of quantitation by the protein 200 plus assay (Table 4), shown by deviations from expected values at the 0.5 M NaCl level that were almost double those displayed by the controls. For the Pro260 analysis, t
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