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)

被引:4
作者
Shimada, Takashi [1 ,2 ]
Toyama, Atsuhiko [1 ]
Aoki, Chikage [1 ]
Aoki, Yutaka [1 ]
Tanaka, Koichi [2 ,3 ]
Sato, Taka-Aki [1 ,2 ]
机构
[1] Shimadzu Co Ltd, Life Sci Res Ctr, Chiyoda Ku, Tokyo 1018448, Japan
[2] Shimadzu Co Ltd, Koichi Tanaka Lab Adv Sci & Technol, Nakagyo Ku, Kyoto 6048511, Japan
[3] Shimadzu Co Ltd, Koichi Tanaka Mass Spectrometry Res Lab, Nakagyo Ku, Kyoto 6048511, Japan
基金
日本学术振兴会;
关键词
ALZHEIMERS-DISEASE; PROTEOMICS; PROTEIN; DEMENTIA; RISK;
D O I
10.1002/rcm.5259
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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, demon-strating the powerful tool of iMS as a biomarker detection method. Copyright (C) 2011 John Wiley & Sons, Ltd.
引用
收藏
页码:3521 / 3526
页数:6
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