ANYL 236 |
| All column separation techniques face the challenge of variations in migration times. Although the non-porous reversed phase columns for protein separation are very reproducible it is not uncommon to see minor changes in migration times for proteins with the same hydrophobicity from different samples. These minor variations may result in complex patterns in allocating the proteins in a data base with migration time as a coordinate. We propose to use a two-marker method to standardize hydrophobicity normally reflected by migration times. One marker is chosen to be extremely hydrophilic while the other is extremely hydrophobic, such that all proteins can be expected to elute between the two markers. Instead of plotting chromatograms using the absorption signal vs. the retention time, the x-axis is plotted in terms of a normalized hydrophobicity index. The advantages of this method will be demonstrated with two dimensional fractionation of plasma proteins using PF-2D system, in conjunction with mass spectrometry protein identification. |
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New Directions and New Techniques in Separation Science and Biomarker Discovery
1:30 PM-4:50 PM, Tuesday, 12 September 2006 Moscone Center -- Room 130, Oral
Division of Analytical Chemistry |