Highly specific label-free protein detection from lysed cells using internally referenced microcantilever sensors

被引:38
作者
Shu, Wenmiao [1 ]
Laurenson, Sophie [2 ]
Knowles, Tuomas P. J. [1 ]
Ferrigno, Paul Ko [2 ]
Seshia, Ashwin A. [1 ,3 ]
机构
[1] Univ Cambridge, Nanosci Ctr, Cambridge CB3 0FF, England
[2] Hutchinson MRC Res Ctr, MRC Canc Cell Unit, Cambridge CB2 2XZ, England
[3] Univ Cambridge, Dept Engn, Cambridge CB2 1PZ, England
基金
英国医学研究理事会; 英国工程与自然科学研究理事会;
关键词
microcantilevers; peptide aptamers; protein fishing;
D O I
10.1016/j.bios.2008.03.036
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
We report the investigation of label-free protein detection directly from lysed cells using microcantilever sensors. The integration of an internally referenced microcantilever sensor combined with peptide aptamer technology enables scalable and label-free detection of proteins from a complex biological environment (e.g. cell lysate). The internally referenced microcantilever sensor was found to be effective in minimizing both the effects of thermal drift and non-specific binding interactions with the backside of the cantilever, thereby allowing protein detection in a complex biological background. Highly specific peptide aptamers are used to modify the cantilever surface to specifically detect less than 80 nM CDK2 protein from yeast cell lysate. This binding of CDK2 on the microcantilever generates a tensile surface stress of average magnitude equal to 70 +/- 22 mN/m. Similar experiments conducted with quartz crystal microbalance (QCM) technology are consistent with the response observed using microcantilever sensors. (C) 2008 Elsevier B.V. All rights reserved.
引用
收藏
页码:233 / 237
页数:5
相关论文
共 25 条
[21]   Design and validation of a neutral protein scaffold for the presentation of peptide aptamers [J].
Woodman, R ;
Yeh, JTH ;
Laurenson, S ;
Ferrigno, PK .
JOURNAL OF MOLECULAR BIOLOGY, 2005, 352 (05) :1118-1133
[22]   Bioassay of prostate-specific antigen (PSA) using microcantilevers [J].
Wu, GH ;
Datar, RH ;
Hansen, KM ;
Thundat, T ;
Cote, RJ ;
Majumdar, A .
NATURE BIOTECHNOLOGY, 2001, 19 (09) :856-860
[23]   Directed evolution of high-affinity antibody mimics using mRNA display [J].
Xu, LH ;
Aha, P ;
Gu, K ;
Kuimelis, RG ;
Kurz, M ;
Lam, T ;
Lim, AC ;
Liu, HX ;
Lohse, PA ;
Sun, L ;
Weng, S ;
Wagner, RW ;
Lipovsek, D .
CHEMISTRY & BIOLOGY, 2002, 9 (08) :933-942
[24]   Rapid and label-free nanomechanical detection of biomarker transcripts in human RNA [J].
Zhang, J. ;
Lang, H. P. ;
Huber, F. ;
Bietsch, A. ;
Grange, W. ;
Certa, U. ;
McKendry, R. ;
Guentgerodt, H. -J. ;
Hegner, M. ;
Gerber, Ch. .
NATURE NANOTECHNOLOGY, 2006, 1 (03) :214-220
[25]   Highly reversible and multi-stage cantilever actuation driven by polyelectrolyte brushes [J].
Zhou, F ;
Shu, WM ;
Welland, ME ;
Huck, WTS .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2006, 128 (16) :5326-5327