Wu Jiaqi, Huang Tiemin
Convergent Bioscience Ltd., Toronto, Ontario, Canada.
Electrophoresis. 2006 Sep;27(18):3584-90. doi: 10.1002/elps.200500889.
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.
在CIEF分析中,样品峰可通过其相对峰位置(RPP)来识别,该相对峰位置仅使用两个内部pI标记物来确定。两个内部pI标记物峰应尽可能紧密地包围样品峰。然后使用两个pI标记物的pI值、峰位置以及样品的峰位置来计算样品峰的RPP值。使用该方法可有效补偿由于盐类导致的pH梯度扭曲。此外,如本文结果所示,无论给定载体两性电解质建立的pH梯度的线性如何,只要使用相同的载体两性电解质运行样品并将盐浓度保持在0 - 15 mM范围内,样品峰在RPP中均可在0.1 pH单位的标准差内(<2%相对标准偏差)被识别。