Optimization of a MALDI TOF-TOF mass spectrometer for intact protein analysis

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Abstract

A MALDI TOF-TOF instrument was optimized and evaluated for intact protein analysis by tandem mass spectrometry. Ion source voltages and delay times were adjusted to affect an up to a 10-fold improvement in fragment ion yield compared to data obtained using default settings employed in peptide analysis. For large peptides (3-4.5 kDa), up to 90% of all possible b- and y-fragment ions were observed, which provides sufficient information for de novo sequencing and unambiguous protein identification. Product ion signals associated with preferential cleavages C-terminal to aspartic acid and glutamic acid residues and N-terminal to proline residues became dominant with increased protein molecular weight. Matrix effects were also evaluated and, among the eight matrices examined, α-cyano-4-hydroxycinnamic acid (CHCA) was found to produce the best intact protein tandem mass spectra for proteins up to 12 kDa. Optimized performance yielded detection limits of 50-125 fmol for proteins of 4 and 12 kDa, respectively. This improved performance has yielded an instrument with potential to be a useful tool in proteomic investigations via analysis of intact proteins. © 2005 American Society for Mass Spectrometry.

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APA

Liu, Z., & Schey, K. L. (2005). Optimization of a MALDI TOF-TOF mass spectrometer for intact protein analysis. Journal of the American Society for Mass Spectrometry, 16(4), 482–490. https://doi.org/10.1016/j.jasms.2004.12.018

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