Peak identification in capillary isoelectric focusing using the concept of relative peak position as determined by two isoelectric point markers.
In CIEF analysis, sample peaks can be identified by their relative peak positions (RPP) that are determined using only two internal pI markers. The two internal pI marker peaks should bracket, as close as possible, the sample peaks. The RPP values of the sample peaks are then calculated using the pI values, peak positions of the two pI markers, and peak position of the sample. Use of this method can effectively compensate for pH gradient distortions that often occur as a result of salts. Also, as shown by the results of this paper, regardless of the linearity of the pH gradient established by the given carrier ampholytes, sample peaks can be identified within an SD of 0.1 pH unit in RPP (<2% RSD) as long as the sample is run using the same carrier ampholytes and maintaining salt concentrations in the range of 0-15 mM.
Wu
Jiaqi
J
Convergent Bioscience Ltd., Toronto, Ontario, Canada. wuj@convergentbiosci.com
Huang
Tiemin
T
D000665:Ampholyte Mixtures / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000592:standards*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000592:standards*; D007526:Isoelectric Point; D011506:Proteins / Q000032:analysis*; D012492:Salts / Q000737:chemistry
D000665:Ampholyte Mixtures; D011506:Proteins; D012492:Salts