Label-free, electrical detection of the SARS virus N-protein with nanowire biosensors utilizing antibody mimics as capture probes.

Antibody mimic proteins (AMPs) are polypeptides that bind to their target analytes with high affinity and specificity, just like conventional antibodies, but are much smaller in size (2-5 nm, less than 10 kDa). In this report, we describe the first application of AMP in the field of nanobiosensors. In(2)O(3) nanowire based biosensors have been configured with an AMP (Fibronectin, Fn) to detect nucleocapsid (N) protein, a biomarker for severe acute respiratory syndrome (SARS). Using these devices, N protein was detected at subnanomolar concentration in the presence of 44 microM bovine serum albumin as a background. Furthermore, the binding constant of the AMP to Fn was determined from the concentration dependence of the response of our biosensors.

[1]  Cees Dekker,et al.  Identifying the mechanism of biosensing with carbon nanotube transistors. , 2008, Nano letters.

[2]  Rui Zhang,et al.  Real-Time, Label-Free Detection of Biological Entities Using Nanowire-Based FETs , 2008, IEEE Transactions on Nanotechnology.

[3]  J. Heath,et al.  A non-oxidative approach toward chemically and electrochemically functionalizing Si(111). , 2006, Journal of the American Chemical Society.

[4]  M. Mrksich,et al.  Using electroactive substrates to pattern the attachment of two different cell populations , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Hans Wolf,et al.  An aptamer-based protein biochip. , 2005, Analytical chemistry.

[6]  Leroy Hood,et al.  Electrochemically programmed, spatially selective biofunctionalization of silicon wires. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[7]  James R Heath,et al.  Quantitative real-time measurements of DNA hybridization with alkylated nonoxidized silicon nanowires in electrolyte solution. , 2006, Journal of the American Chemical Society.

[8]  M. Lai,et al.  The Molecular Biology of Coronaviruses , 1997, Advances in Virus Research.

[9]  Muhammad A. Alam,et al.  Screening-limited response of nanobiosensors. , 2007, Nano letters.

[10]  A. Star,et al.  Carbon Nanotube Field‐Effect‐Transistor‐Based Biosensors , 2007 .

[11]  Mark A. Reed,et al.  Label-free immunodetection with CMOS-compatible semiconducting nanowires , 2007, Nature.

[12]  Tamar Frankel [The theory and the practice...]. , 2001, Tijdschrift voor diergeneeskunde.

[13]  Xuguang Li,et al.  SARS coronavirus: Unusual lability of the nucleocapsid protein , 2008, Biochemical and Biophysical Research Communications.

[14]  Kang L. Wang,et al.  One-dimensional transport of In2O3 nanowires , 2005 .

[15]  Chao Li,et al.  Complementary detection of prostate-specific antigen using In2O3 nanowires and carbon nanotubes. , 2005, Journal of the American Chemical Society.

[16]  Marek Kosmulski,et al.  Pristine Points of Zero Charge of Gallium and Indium Oxides. , 2001, Journal of colloid and interface science.

[17]  F. Taguchi [Molecular biology of coronaviruses]. , 1990, Uirusu.

[18]  G. Gruner Carbon nanotube transistors for biosensing applications , 2006 .

[19]  C. Lieber,et al.  Nanowire Nanosensors for Highly Sensitive and Selective Detection of Biological and Chemical Species , 2001, Science.

[20]  L. Babiuk,et al.  Severe acute respiratory syndrome coronavirus nucleocapsid protein expressed by an adenovirus vector is phosphorylated and immunogenic in mice. , 2005, The Journal of general virology.

[21]  Fred J Sigworth,et al.  Importance of the Debye screening length on nanowire field effect transistor sensors. , 2007, Nano letters.

[22]  C. Li,et al.  Selective functionalization of In2O3 nanowire mat devices for biosensing applications. , 2005, Journal of the American Chemical Society.

[23]  C. Zhou,et al.  Tuning electronic properties of In2O3 nanowires by doping control , 2004 .

[24]  Paul L. McEuen,et al.  High Performance Electrolyte Gated Carbon Nanotube Transistors , 2002 .

[25]  A. Plückthun,et al.  Engineering novel binding proteins from nonimmunoglobulin domains , 2005, Nature Biotechnology.

[26]  K. Struhl,et al.  Current Protocols in Molecular Biology (New York: Greene Publishing Associates and Wiley-Interscience). Host-Range Shuttle System for Gene Insertion into the Chromosomes of Gram-negative Bacteria. , 1988 .

[27]  C. Zhou,et al.  Synthesis, Electronic Properties, and Applications of Indium Oxide Nanowires , 2003, Annals of the New York Academy of Sciences.

[28]  Andreas Plückthun,et al.  Engineered proteins as specific binding reagents. , 2005, Current opinion in biotechnology.

[29]  Y. Chang,et al.  Carbon nanotube DNA sensor and sensing mechanism. , 2006, Nano letters.

[30]  Qian Wang,et al.  An investigation of the mechanisms of electronic sensing of protein adsorption on carbon nanotube devices. , 2004, Journal of the American Chemical Society.

[31]  Irving Langmuir,et al.  The constitution and fundamental properties of solids and liquids. Part II.—Liquids , 1917 .

[32]  Gengfeng Zheng,et al.  Nanowire sensors for medicine and the life sciences. , 2006, Nanomedicine.

[33]  J. Crowther ELISA. Theory and practice. , 1995, Methods in molecular biology.

[34]  Arnaud Buhot,et al.  On the hybridization isotherms of DNA microarrays: the Langmuir model and its extensions , 2006 .