Designed Nano/Micro-Reactors For Mass Spectrometry Based Anlysis Of Membrane Proteins

Compared with traditional nanoparticles in versatile applications,the controlled pore size and surface chemistry modification of nanoporous materials make them quite popular in a number of research fields including biomedical science.In particular,due to their high pore volume,large pore size and controlled surface modification,the macroporous silica materials(pore size >50nm)have many advantages over other materials in biomolecule detection(e.g.,proteins).Mass spectrometry(MS)-based proteome profiling is essential for molecular diagnostics in modern biomedical study.To date,sample preparation including protein extraction and proteolysis is still very challenging and lack of efficiency.In this article,we have prepared the macroporous siliceous foams(MSF)with large pore size of ~70 nm and hydrophobic surface(contact angle over 100°).The modified MSF was employed for selective extraction and in situ tryptic digestion of membrane proteins coupled with MS based profiling in biological samples.The optimized MSF was selected in detection of a standard membrane protein(bacteriorhodopsin)and further applied to Hela cell lysates yielding 112 membrane proteins identified.Moreover,when integrated with micropipette tips,the nano-micro-reactors were demonstrated as high performance devices for fast(~minutes)and multiplexing proteolysis to assist the profiling of membrane proteome.With the enhanced performance,our work built a novel,efficient and miniaturized platform for facile proteomic sample preparation,which is promising for advanced biomarkers discovery in biomedical study.