Study of Derivatization of Peptides and Proteins for Fluorescence Detection Using Capillary Electrophoresis and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry

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Authors

VRÁBEL Patrik PREISLER Jan RYVOLOVÁ Markéta ŘEHULKOVÁ Helena TÁBORSKÝ Petr

Year of publication 2005
Type Conference abstract
MU Faculty or unit

Faculty of Science

Citation
Description We presented an effective combination of two analytical techniques (CE-MALDI MS) for sensitive identification of fluorescently labeled polypeptides and optimization of derivatization procedure. In analysis of compounds, which included derivatization step, the detection was found to be limited by chemistry and separation rather than by the instrument performance. This was confirmed on CE-LIF of peptides and proteins derivatized with rhodamine B isothiocyanate (RBITC). Identification of all derivatized species was not possible with fluorescence detection; therefore CE experiments with mass spectrometric detection of derivatization products were applied. Optimized CE separation of peptides and small proteins was coupled to MALDI MS; common MALDI matrices were used. This experimental work confirmed suitability of MALDI MS for detection and identification of peptides and proteins derivatized with rhodamine-based dyes. The information from the experiment will be used for optimizing of the derivatization step.
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