Limitation of immunoaffinity column for the removal of abundant proteins from plasma in quantitative plasma proteomics

作者：Ichibangase Tomoko; Moriya Kyoji; Koike Kazuhiko; Imai Kazuhiro^*

来源：Biomedical Chromatography, 2009, 23(5): 480-487.

DOI：10.1002/bmc.1139

• plasma
• proteomics
• immunoaffinity column
• abundant protein
• FD-LC-MS/MS method

In plasma proteomics, before a proteome analysis, it is essential to prepare protein samples without high-abundance proteins, including albumin, via specific preparation techniques, such as immunoaffinity capture. However, our preliminary experiments suggested that functional changes with use after the ability of the immunoaffinity column. Thus, in this study, to evaluate the changes of the removal ability of abundant proteins from plasma by the immunoaffinity column, plasma proteome analysis was performed for the long-term test for the reproducibility of the affinity column using the fluorogenic derivatization-liquid chromatography-tandem mass spectrometry method combined with an IgY column. The specific adsorption for albumin decreased with an increase in the number of the column usage before its expiration date. Moreover, it was demonstrated that hydrophobic high molecular weight compounds in plasma adsorbed onto the column materials surface contributed to the functional changes from specific immunoaffinity adsorption into hydrophobic interaction. These results suggested that, in quantitative plasma proteomics studies, it is important to keep in mind the risk of not only the nonselective loss but also the changes in the adsorption ability of the immunoafinity column.

• 出版日期2009-5