A hairpin aptamer-based electrochemical biosensing platform for the sensitive detection of proteins.

An aptamer-based electrochemical sensing platform for the direct protein detection has been developed using IgE and a specifically designed aptamer with hairpin structure as the model analyte and probe sequence, respectively. In the absence of IgE, the aptamer immobilized on an electrode surface forms a large hairpin due to the hybridization of the two complementary arm sequences, and peak currents of redox species dissolved in solution can be achieved. However, the target protein binding can not only cause the increase of the dielectric layer but also trigger the significant conformational switching of the aptamer due to the opening of the designed hairpin structure that pushes the biomolecule layer/electrolyte interface away from the electrode surface, suppressing substantially the electron transfer (eT) and resulting in a strong detection signal. The detection limit of 3.6x10(-11)M and linear response range of 5.4x10(-11) to 3.6x10(-8)M are achieved without any amplifier. The selectivity is confirmed by interference test. More importantly, an innovative concept of adapting intelligently a surface-confined aptamer sequence is introduced, and the limitations of the conventional electrochemical aptasensors have been overcome. The proposed sensing scheme is expected to become a promising strategy for the detection of proteins and other biomacromolecules.

[1]  X. Cui,et al.  Reagentless aptamer based impedance biosensor for monitoring a neuro-inflammatory cytokine PDGF. , 2007, Biosensors & bioelectronics.

[2]  J. Kinet,et al.  High-affinity oligonucleotide ligands to human IgE inhibit binding to Fc epsilon receptor I. , 1996, Journal of immunology.

[3]  Ciara K O'Sullivan,et al.  Reusable impedimetric aptasensor. , 2005, Analytical chemistry.

[4]  Yi Lu,et al.  Adenosine-dependent assembly of aptazyme-functionalized gold nanoparticles and its application as a colorimetric biosensor. , 2004, Analytical chemistry.

[5]  Giridharan Gokulrangan,et al.  DNA aptamer-based bioanalysis of IgE by fluorescence anisotropy. , 2005, Analytical chemistry.

[6]  Kevin W Plaxco,et al.  A reagentless signal-on architecture for electronic, aptamer-based sensors via target-induced strand displacement. , 2005, Journal of the American Chemical Society.

[7]  Kenzo Maehashi,et al.  Label-free protein biosensor based on aptamer-modified carbon nanotube field-effect transistors. , 2007, Analytical chemistry.

[8]  M. Stojanović,et al.  Aptamer-based folding fluorescent sensor for cocaine. , 2001, Journal of the American Chemical Society.

[9]  Milan N Stojanovic,et al.  Fluorescent Sensors Based on Aptamer Self-Assembly. , 2000, Journal of the American Chemical Society.

[10]  Chunhai Fan,et al.  Aptamer-based biosensors , 2008 .

[11]  Yusuke Ohtani,et al.  Bis-pyrene labeled DNA aptamer as an intelligent fluorescent biosensor. , 2003, Bioorganic & medicinal chemistry letters.

[12]  Ciara K O'Sullivan,et al.  Reagentless, reusable, ultrasensitive electrochemical molecular beacon aptasensor. , 2006, Journal of the American Chemical Society.

[13]  Shanlin Pan,et al.  Chemical control of electrode functionalization for detection of DNA hybridization by electrochemical impedance spectroscopy. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[14]  Kevin W Plaxco,et al.  Aptamer-based electrochemical detection of picomolar platelet-derived growth factor directly in blood serum. , 2007, Analytical chemistry.

[15]  Chunli Bai,et al.  Signaling aptamer/protein binding by a molecular light switch complex. , 2004, Analytical chemistry.

[16]  杨朝勇 Light-switching excimer probes for rapid protein monitoring in complex biological fluids , 2005 .

[17]  J. SantaLucia,et al.  A unified view of polymer, dumbbell, and oligonucleotide DNA nearest-neighbor thermodynamics. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[18]  K. Weeks,et al.  Facile conversion of aptamers into sensors using a 2'-ribose-linked fluorophore. , 2005, Journal of the American Chemical Society.

[19]  A. Bard,et al.  Electron transfer at self-assembled monolayers measured by scanning electrochemical microscopy. , 2004, Journal of the American Chemical Society.

[20]  Guo-Li Shen,et al.  Reusable electrochemical sensing platform for highly sensitive detection of small molecules based on structure-switching signaling aptamers. , 2007, Analytical chemistry.

[21]  B. J. Sutton,et al.  The human IgE network , 1993, Nature.

[22]  Hans Wolf,et al.  An aptamer-based quartz crystal protein biosensor. , 2002, Analytical chemistry.

[23]  Neal W. Woodbury,et al.  Exploring the sequence space of a DNA aptamer using microarrays , 2007, Nucleic acids research.

[24]  Ciara K O'Sullivan,et al.  Aptamer conformational switch as sensitive electrochemical biosensor for potassium ion recognition. , 2006, Chemical communications.

[25]  Xiaobo Yu,et al.  Label-free electrochemical detection for aptamer-based array electrodes. , 2005, Analytical chemistry.

[26]  W. Tan,et al.  Molecular aptamer for real-time oncoprotein platelet-derived growth factor monitoring by fluorescence anisotropy. , 2001, Analytical chemistry.

[27]  G. Shen,et al.  Reversible electronic nanoswitch based on DNA G-quadruplex conformation: a platform for single-step, reagentless potassium detection. , 2008, Biomaterials.

[28]  A. Heeger,et al.  An electronic, aptamer-based small-molecule sensor for the rapid, label-free detection of cocaine in adulterated samples and biological fluids. , 2006, Journal of the American Chemical Society.

[29]  A. Steel,et al.  Electrochemical quantitation of DNA immobilized on gold. , 1998, Analytical chemistry.

[30]  Juewen Liu,et al.  Fast colorimetric sensing of adenosine and cocaine based on a general sensor design involving aptamers and nanoparticles. , 2005, Angewandte Chemie.

[31]  Chunhai Fan,et al.  A target-responsive electrochemical aptamer switch (TREAS) for reagentless detection of nanomolar ATP. , 2007, Journal of the American Chemical Society.

[32]  Yingfu Li,et al.  Structure-switching signaling aptamers. , 2003, Journal of the American Chemical Society.

[33]  R. Kennedy,et al.  Aptamers as ligands in affinity probe capillary electrophoresis. , 1998, Analytical chemistry.

[34]  Michael Zuker,et al.  Mfold web server for nucleic acid folding and hybridization prediction , 2003, Nucleic Acids Res..

[35]  Ping Yu,et al.  Aptamer-based electrochemical sensors with aptamer-complementary DNA oligonucleotides as probe. , 2008, Analytical chemistry.