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Improved sensitivity for insulin in matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry by premixing α ‐cyano‐4‐hydroxycinnamic acid matrix with transferrin
Author(s) -
Kobayashi Tetsu,
Kawai Hiroshi,
Suzuki Takuo,
Kawanishi Toru,
Hayakawa Takao
Publication year - 2004
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1465
Subject(s) - chemistry , mass spectrometry , matrix assisted laser desorption/ionization , chromatography , bovine serum albumin , time of flight mass spectrometry , detection limit , transferrin , desorption , biochemistry , ionization , ion , organic chemistry , adsorption
This report describes an enhancement of the signal intensities of proteins and peptides in matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS). When α ‐cyano‐4‐hydroxycinnamic acid (CHCA) premixed with human transferrin (Tf) was used as a matrix, the signal intensity of insulin was amplified to more than ten times that of the respective control in CHCA without Tf. The detection limit of insulin was 0.39 fmol on‐probe in the presence of Tf, while it was 6.3 fmol in the absence of Tf. The signal intensity of insulin was also enhanced when the CHCA matrix was premixed with proteins other than Tf (80 kDa), such as horse ferritin (20 kDa), bovine serum albumin (BSA, 66 kDa), or human immunoglobulin G (150 kDa). The optimum spectrum of insulin was obtained when the added amount of protein was in the range 0.26–0.62 pmol, regardless of the molecular weight of the added protein. Tf and BSA outperformed the other tested proteins, as determined by improvements in the resulting spectra. When the mass spectra of several peptides and proteins were recorded in the presence of Tf or BSA, the signal intensities of large peptides such as glucagon were enhanced, though those of smaller peptides were not enhanced. In addition, the signal enhancement achieved with Tf and BSA was more pronounced for the proteins, including cytochrome C, than for the large peptides. This enhancement effect could be applied to improve the sensitivity of MALDI‐TOFMS to large peptides and proteins. Copyright © 2004 John Wiley & Sons, Ltd.