Proteomic profiling of platelet proteins by trypsin immobilized self-assembled monolayers digestion chip and protein identification using electrospray ionization tandem mass spectrometry

Journal of Biomedical Materials Research. Part a
Yu-Chang TyanMichael Grunze

Abstract

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 di...Continue Reading

References

Nov 1, 1991·Journal of Biomedical Materials Research·Y ItoY Imanishi
Aug 1, 1993·Biomaterials·I K KangH B Lee
May 4, 1992·Physical Review Letters·S A JoyceR M Crooks
Jun 20, 1996·Chemical Reviews·Abraham Ulman
May 29, 2002·Thrombosis Research·Tze-Wen ChungShu-Hsun Chu
Sep 17, 2002·Briefings in Bioinformatics·Claire O'DonovanRolf Apweiler
Nov 13, 2003·Journal of Biomedical Materials Research. Part a·Yu-Chang TyanChia-Chieh Chen

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Citations

May 26, 2006·Analytical and Bioanalytical Chemistry·Lee Wah LimToyohide Takeuchi
Aug 24, 2006·Blood·Dmitri V GnatenkoWadie F Bahou
Mar 8, 2005·Journal of Mass Spectrometry : JMS
May 23, 2007·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Thomas ThieleAndreas Greinacher

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