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Proteomic profiling of platelet proteins by trypsin immobilized self‐assembled monolayers digestion chip and protein identification using electrospray ionization tandem mass spectrometry
Author(s) -
Tyan YuChang,
Liao JiunnDer,
Jong ShiangBin,
Liao PaoChi,
Yang MingHui,
Chang YinWei,
Klauser Ruth,
Himmelhaus Michael,
Grunze Michael
Publication year - 2004
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.30129
Subject(s) - electrospray ionization , self assembled monolayer , trypsin , monolayer , biomolecule , mass spectrometry , chemistry , protein mass spectrometry , x ray photoelectron spectroscopy , amide , attenuated total reflection , covalent bond , tandem mass spectrometry , analytical chemistry (journal) , fourier transform infrared spectroscopy , materials science , chromatography , infrared spectroscopy , organic chemistry , chemical engineering , enzyme , biochemistry , engineering
Self‐assembled monolayers (SAMs) on coinage metal provide versatile modeling systems for studies of interfacial electron transfer, biological interactions, molecular recognition, and other interfacial phenomena. Recently, the bonding of enzyme to SAMs of alkanethiols onto Au electrode surfaces was exploited to produce a bio‐sensing system. In this work, the attachment of trypsin to a SAMs surface of 11‐mercaptoundecanoic acid was achieved using water soluble 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide and N‐hydroxysulfosuccinimide as coupling agent. Experimental results have revealed that the X‐ray Photoelectron Spectroscopy (XPS) C1s core levels at 286.3 and 286.5 eV (C with N), 288.1 eV (amide bond), and 289.3 eV (carboxyl) illustrate the immobilization of trypsin. These data were also in good agreement with Fourier‐Transformed Infrared Reflection‐Attenuated Total Reflection (FTIR‐ATR) spectra for the peaks valued at 1659.4 cm −1 (amide I) and 1546.6 cm −1 (amide II). Using electrospray ionization tandem mass spectrometry (ESI‐MS/MS) observations, analytical results have demonstrated the platelet proteins digestion of the immobilized trypsin on the functionalized SAMs surface. For such surfaces, platelet proteins were digested on the trypsin‐immobilized SAMs surface, which shows the enzyme digestion ability of the immobilized trypsin. The terminal groups of the SAMs structure can be further functionalized with biomolecules or antibodies to develop surface‐base diagnostics, biosensors, or biomaterials. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res 71A: 90–97, 2004