A novel PVDF membrane for on-membrane identification of gel-resolved proteins by mass spectrometry

在20世纪90年代，人们开发了将蛋白质从凝胶转移到PVDF膜上的技术，并用MALDI-MS鉴定膜上的蛋白质。然而，固定在膜上的多肽和蛋白质的低电离效率往往使这些技术毫无用处。因此，开发能够有效电离多肽和蛋白质的膜用于MS分析是非常必要的。在本研究中，我们发现0.1-μm孔聚偏氟乙烯膜可用于高效的电印迹和MALDI-MS分析。与传统的PVDF膜相比，该膜在多肽的MALDI-MS分析中表现出较高的离子产率。用2-DE分离了38种具有不同理化性质的酵母蛋白，并将其固定在0.1-μm孔的聚偏氟乙烯膜上，用MALDI-MS/MS成功地鉴定了这些蛋白质。

In the 1990s, techniques were developed to transfer proteins from gels onto PVDF membranes, and to identify the proteins on the membranes by MALDI-MS. However, the low ionization efficiency of peptides and proteins immobilized on the membranes often renders these techniques useless. Therefore, it is essential to develop membranes on which peptides and proteins can be efficiently ionized for MS analysis. In this study, we found that 0.1-μm pores PVDF membranes can be used for highly efficient and effective electroblotting and MALDI-MS analysis. This membrane showed high ion yield in MALDI-MS analysis of peptides compared to conventional PVDF membranes. In all, 38 yeast proteins with various physicochemical characteristics were separated by 2-DE, immobilized onto the 0.1-μm pore PVDF membranes, and successfully identified by MALDI-MS/MS.