Indirect ultrasonication for protein quantification and peptide mass mapping through mass spectrometry-based techniques |
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Authors: | R.J. Carreira M. Reboiro-Jato F. Fdez-Riverola |
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Affiliation: | a REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Quinta da Torre, 2829-516 Monte de Caparica, Portugal b Department of Physical Chemistry, Faculty of Sciences at Ourense Campus, University of Vigo at Ourense Campus, E-32004 Ourense, Spain c SING Group, Informatics Department, Higher Technical School of Computer Engineering, University of Vigo, Ourense, Spain |
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Abstract: | We report in this work a fast protocol for protein quantification and for peptide mass mapping that rely on 18O isotopic labeling through the decoupling procedure. It is demonstrated that the purity and source of trypsin do not compromise the labeling degree and efficiency of the decoupled labeling reaction, and that the pH of the labeling reaction is a critical factor to obtain a significant 18O double labeling. We also show that the same calibration curve can be used for MALDI protein quantification during several days maintaining a reasonable accuracy, thus simplifying the handling of the quantification process. In addition we demonstrate that 18O isotopic labeling through the decoupling procedure can be successfully used to elaborate peptide mass maps. BSA was successfully quantified using the same calibration curve in different days and plasma from a freshwater fish, Cyprinus carpio, was used to elaborate the peptide mass maps. |
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Keywords: | 18O-labeling Ultrasound Sonoreactor Peptide mass mapping Differential protein expression MALDI-TOF-MS Protein quantification |
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