Limitation of immunoaffinity column for the removal of abundant proteins from plasma in quantitative plasma proteomics.
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 alter 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.
Ichibangase
Tomoko
T
Research Institute of Pharmaceutical Sciences, Musashino University, Tokyo, Japan.
Moriya
Kyoji
K
Koike
Kazuhiko
K
Imai
Kazuhiro
K
D000818:Animals; D001798:Blood Proteins / Q000032:analysis* / Q000276:immunology; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D006801:Humans; D007136:Immunoglobulins / Q000276:immunology; D007163:Immunosorbent Techniques / Q000295:instrumentation*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008855:Microscopy, Electron, Scanning; D020543:Proteome / Q000032:analysis; D040901:Proteomics / Q000295:instrumentation* / Q000379:methods*; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000276:immunology / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013499:Surface Properties; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods
D001798:Blood Proteins; C016512:IgY; D007136:Immunoglobulins; D020543:Proteome; D012709:Serum Albumin