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Fully automated chip-based nanoelectrospray combined with electron transfer dissociation for high throughput top-down proteomics

Fully automated chip-based nanoelectrospray combined with electron transfer dissociation for high throughput top-down proteomics

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FLANGEA, Corina, Catalin SCHIOPU, Florina CAPITAN, Cristina MOSOARCA, Marilena MANEA, Eugen SISU, Alina D. ZAMFIR, 2012. Fully automated chip-based nanoelectrospray combined with electron transfer dissociation for high throughput top-down proteomics. In: Central European Journal of Chemistry. 11(1), pp. 25-34. ISSN 1895-1066. eISSN 1644-3624

@article{Flangea2012Fully-21148, title={Fully automated chip-based nanoelectrospray combined with electron transfer dissociation for high throughput top-down proteomics}, year={2012}, doi={10.2478/s11532-012-0130-2}, number={1}, volume={11}, issn={1895-1066}, journal={Central European Journal of Chemistry}, pages={25--34}, author={Flangea, Corina and Schiopu, Catalin and Capitan, Florina and Mosoarca, Cristina and Manea, Marilena and Sisu, Eugen and Zamfir, Alina D.} }

Manea, Marilena Sisu, Eugen 2013-01-10T08:53:03Z Fully automated chip-based nanoelectrospray combined with electron transfer dissociation for high throughput top-down proteomics Zamfir, Alina D. The conventional protocol for protein identification by electrospray ionization mass spectrometry (MS) is based on enzymatic digestion which renders peptides to be analyzed by liquid chromatography-MS and collision-induced dissociation (CID) multistage MS, in the so-called bottom-up approach. Though this method has brought a significant progress to the field, many limitations, among which, the low throughput and impossibility to characterize in detail posttranslational modifications in terms of site(s) and structure, were reported. Therefore, the research is presently focused on the development of procedures for efficient top-down fragmentation of intact protein ions. In this context, we developed here an approach combining fully automated chip-based-nanoelectrospray ionisatio (nanoESI), performed on a NanoMate robot, with electron transfer dissociation (ETD) for peptide and top-down protein sequencing and identification. This advanced analytical platform, integrating robotics, microfluidics technology, ETD and alternate ETD/CID, was tested<br />and found ideally suitable for structural investigation of peptides and modified/functionalized peptides as well as for top-down analysis of medium size proteins by tandem MS experiments of significantly increased throughput and sensitivity. The obtained results indicate<br />that NanoMate-ETD and ETD/CID may represent a viable alternative to the current MS strategies, with potential to develop into a method of routine use for high throughput top-down proteomics. Schiopu, Catalin 2013-01-10T08:53:03Z Capitan, Florina Zamfir, Alina D. Manea, Marilena Mosoarca, Cristina Capitan, Florina Flangea, Corina Schiopu, Catalin Mosoarca, Cristina eng 2012 Flangea, Corina Central European Journal of Chemistry ; 11 (2013), 1. - S. 25-34 deposit-license Sisu, Eugen

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