Differential Denaturation of Serum Proteome Reveals a Significant Amount of Hidden Information in Complex Mixtures of Proteins

摘要: Recently developed proteomic technologies allow to profile thousands of proteins within a high-throughput approach towards biomarker discovery, although results are not as satisfactory expected. In the present study we demonstrate that serum proteome denaturation is key underestimated feature; in fact, new differential protocol better discriminates according their electrophoretic mobility compared single-denaturation protocols. Sixty nine different treatments were tested and 3 most discriminating ones selected (TRIDENT analysis) applied human sera, showing significant improvement protein discrimination confirmed by MALDI-TOF/MS LC-MS/MS identification, depending on type applied. Thereafter sera from mice patients carrying cutaneous melanoma analyzed through TRIDENT. Nine 8 bands found differentially expressed healthy controls (p<0.05); three them found, for first time, significantly modulated: α2macroglobulin (down-regulated melanoma, p<0.001), Apolipoprotein-E Apolipoprotein-A1 (both up-regulated p<0.04), both humans. The modulation was immunological methods. Other less abundant (e.g. gelsolin) modulated (p<0.05). Conclusions: i) contains large amount information, still neglected, related folding; ii) careful may improve analytical procedures involving complex mixtures; iii) highlights interesting differences between cancer sera.