Detection of adherent cells using electrochemical impedance spectroscopy based on molecular recognition of integrin β1

Abstract Mouse anti-human integrin β1 monoclonal antibody was assembled on glass carbon electrode by layer-by-layer adsorption. The determination of cervical cancer HeLa cells was performed using electrochemical impedance spectroscopy based on the immunoreaction between integrin β1 on cell membrane and the antibody immobilized on the electrode-surface. The factors influencing cell-detection, such as the concentration of gold nanoparticles in the modified film, the drying time of the modified film, the concentration of antibody and the binding time of antibody, were investigated, respectively. Under optimum conditions, the increasing extent of electrode-transfer resistance depended linearly on the cell concentration in the range of 1.0 × 104 to 2.0 × 106 cells mL−1 with a detection limit of 3.5 × 103 cells mL−1.

[1]  M. Shindel,et al.  Molecular self-assembly of solid-supported protein crystals. , 2008, Langmuir : the ACS journal of surfaces and colloids.

[2]  Yi Wang,et al.  A 3D-impedimetric immunosensor based on foam Ni for detection of sulfate-reducing bacteria , 2010 .

[3]  N. Pieczonka,et al.  Gold nanoparticle embedded, self-sustained chitosan films as substrates for surface-enhanced Raman scattering. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[4]  Richard O. Hynes,et al.  Integrins: Versatility, modulation, and signaling in cell adhesion , 1992, Cell.

[5]  C. Mao,et al.  Surface-mediated DNA self-assembly. , 2009, Journal of the American Chemical Society.

[6]  A. Malik,et al.  Electrical method for detection of endothelial cell shape change in real time: assessment of endothelial barrier function. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[7]  Y. Ci,et al.  Voltammetric behavior of mammalian tumor cells and bioanalytical applications in cell metabolism. , 1999, Bioelectrochemistry and bioenergetics.

[8]  Michael J. Natan,et al.  Kinetic Control of Interparticle Spacing in Au Colloid-Based Surfaces: Rational Nanometer-Scale Architecture , 1996 .

[9]  Aiguo Wu,et al.  A method to construct a third-generation horseradish peroxidase biosensor: self-assembling gold nanoparticles to three-dimensional sol-gel network. , 2002, Analytical chemistry.

[10]  Qingjun Liu,et al.  Impedance studies of bio-behavior and chemosensitivity of cancer cells by micro-electrode arrays. , 2009, Biosensors & bioelectronics.

[11]  Hidenori Suzuki,et al.  The SPR signal in living cells reflects changes other than the area of adhesion and the formation of cell constructions. , 2007, Biosensors & bioelectronics.

[12]  Tit Meng Lim,et al.  Detection of Saccharomyces cerevisiae immobilized on self-assembled monolayer (SAM) of alkanethiolate using electrochemical impedance spectroscopy , 2005 .

[13]  Yanbin Li,et al.  Interdigitated Array microelectrode-based electrochemical impedance immunosensor for detection of Escherichia coli O157:H7. , 2004, Analytical chemistry.

[14]  Feng Yan,et al.  A self-assembled monolayer based electrochemical immunosensor for detection of leukemia K562A cells , 2007 .

[15]  F. Aoudjit,et al.  Integrin signaling inhibits paclitaxel-induced apoptosis in breast cancer cells , 2001, Oncogene.

[16]  G. Frens.  Controlled Nucleation for the Regulation of the Particle Size in Monodisperse Gold Suspensions , 1973 .

[17]  Y. Ci,et al.  Photoelectric behavior of mammalian cells and its bioanalytical applications , 1998 .

[18]  P. Hammond.  Form and Function in Multilayer Assembly: New Applications at the Nanoscale , 2004 .

[19]  W. Dalton,et al.  Cell Adhesion Mediated Drug Resistance (CAM-DR): Role of Integrins and Resistance to Apoptosis in Human Myeloma Cell Lines , 1999 .

[20]  Y. Lvov,et al.  Cellulose fiber-enzyme composites fabricated through layer-by-layer nanoassembly. , 2007, Biomacromolecules.

[21]  R. G. Freeman,et al.  Preparation and Characterization of Au Colloid Monolayers , 1995 .

[22]  Itamar Willner,et al.  DNAzyme-Functionalized Au Nanoparticles for the Amplified Detection of DNA or Telomerase Activity , 2004 .

[23]  Bernard Lachance,et al.  An in-depth analysis of electric cell-substrate impedance sensing to study the attachment and spreading of mammalian cells. , 2002, Analytical chemistry.

[24]  Shouzhuo Yao,et al.  Monitoring of cell growth and assessment of cytotoxicity using electrochemical impedance spectroscopy. , 2006, Biochimica et biophysica acta.

[25]  Jian-hui Jiang,et al.  An electrochemical impedance immunosensor with signal amplification based on Au-colloid labeled antibody complex , 2006 .

[26]  Ying Zhang,et al.  A reagentless amperometric immunosensor based on gold nanoparticles/thionine/Nafion-membrane-modified gold electrode for determination of α-1-fetoprotein , 2005 .

[27]  Y. Chai,et al.  Electrochemical Immunoanalysis for Carcinoembryonic Antigen Based on Multilayer Architectures of Gold Nanoparticles and Polycation Biomimetic Interface on Glassy Carbon Electrode , 2006 .

[28]  H D Humes,et al.  Structure and function of cell adhesion molecules. , 1999, The American journal of medicine.

[29]  M. Yousaf,et al.  Total internal reflection fluorescence microscopy of cell adhesion on patterned self-assembled monolayers on gold. , 2009, Langmuir : the ACS journal of surfaces and colloids.

[30]  Hsien-Chang Chang,et al.  Electrochemical evaluation of avidin-biotin interaction on self-assembled gold electrodes , 2005 .

[31]  Jia-cong Shen,et al.  Biomacromolecules electrostatic self-assembly on 3-dimensional tissue engineering scaffold. , 2004, Biomacromolecules.

[32]  Z. Su,et al.  Biorecognition-Driven Self-Assembly of Gold Nanorods: A Rapid and Sensitive Approach toward Antibody Sensing , 2007 .

[33]  Tomokazu Yoshimura,et al.  Antioxidant-potentiality of gold-chitosan nanocomposites , 2003 .

[34]  James R. Johnson,et al.  Self-assembly of fluorescent inclusion complexes in competitive media including the interior of living cells. , 2007, Journal of the American Chemical Society.

[35]  G. Whitesides,et al.  Self-assembled monolayers of thiolates on metals as a form of nanotechnology. , 2005, Chemical reviews.

[36]  J. Randles.  Kinetics of rapid electrode reactions , 1947 .

[37]  A. Zebda,et al.  Electrical resistivity dependence of semi-conductive oxide electrode on the label-free electrochemical detection of DNA , 2010 .

[38]  Gero Decher,et al.  Fuzzy Nanoassemblies: Toward Layered Polymeric Multicomposites , 1997 .