Direct antigen detection from immunoprecipitated beads using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; a new method for immunobeads-mass spectrometry (iMS)

Autor:	Chikage Aoki, Yutaka Aoki, Takashi Shimada, Atsuhiko Toyama, Taka-Aki Sato, Koichi Tanaka
Rok vydání:	2011
Předmět:	Detection limit chemistry.chemical_classification Chromatography Protein mass spectrometry Biomolecule Organic Chemistry Peptide Mass spectrometry Analytical Chemistry Surface-enhanced laser desorption/ionization chemistry Desorption Spectroscopy Alkyl
Zdroj:	Rapid Communications in Mass Spectrometry. 25:3521-3526
ISSN:	0951-4198
Popis:	One-step detection of biological molecules is one of the principal techniques for clinical diagnosis, and the potential of mass spectrometry for biomarker detection has been a promising new approach in the field of medical sciences. We demonstrate here a new and high-sensitivity method that we termed immunobeads-mass spectrometry (iMS), which combines conventional immunoprecipitation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The key feature of iMS is the MS-compatible condition of immunoprecipitation using detergents with a monosaccaride-C8 alkyl chain or a disaccharide-C10 alkyl chain, and the minimized number of steps required for high-sensitivity detection of target peptides in serum or biological fluid. This was achieved by optimizing the wash buffer and subjecting the immunobeads directly to MALDI-TOF MS analysis. Using this method, we showed that 1 fmol of amyloid beta peptide spiked in serum was readily detectable, demonstrating the powerful tool of iMS as a biomarker detection method.
Databáze:	OpenAIRE
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