Detection of DNA hybridization by a field‐effect transistor with covalently attached catcher molecules

Electronic DNA sensors based on field-effect transistor (FET) arrays operating in liquid environments offer an alternative method for the detection of biomolecular binding events without the requirement to label the analyte molecules. In order to obtain reproducible signals from such sensors, the attachment of the probe molecules to the gate area of the transistor chips requires a well-controlled supramolecular architecture. The FET chips used in this work are non-metallized, 8-channel transistor devices with micrometer dimensions of the gate structures, which are encapsulated to be dipped into an analyte solution. Two identical chips were functionalized with DNA catcher molecules of different sequences and read out in a differential measurement approach. In this article, we describe the details of the surface modification and covalent attachment of the catcher DNA. The surface modification was characterized by XPS, Fourier transform infrared spectroscopy (FT-IR), fluorescence microscopy and imaging ellipsometry. Electrical FET recordings of the immobilization and hybridization of DNA molecules are presented.

[1]  A. Offenhäusser,et al.  Field-effect transistor array for monitoring electrical activity from mammalian neurons in culture. , 1997, Biosensors & bioelectronics.

[2]  Tomoji Kawai,et al.  Direct Deoxyribonucleic Acid Detection Using Ion-Sensitive Field-Effect Transistors Based on Peptide Nucleic Acid , 2004 .

[3]  Jang-Kyoo Shin,et al.  Detection of DNA and Protein Molecules Using an FET‐Type Biosensor with Gold as a Gate Metal , 2004 .

[4]  Jang-Kyoo Shin,et al.  Field Effect Transistor-based Bimolecular Sensor Employing a Pt Reference Electrode for the Detection of Deoxyribonucleic Acid Sequence , 2004 .

[5]  Yuji Miyahara,et al.  Potentiometric Detection of Single Nucleotide Polymorphism by Using a Genetic Field‐effect transistor , 2005, Chembiochem : a European journal of chemical biology.

[6]  Larry J Kricka,et al.  Stains, labels and detection strategies for nucleic acids assays , 2002, Annals of clinical biochemistry.

[7]  L. Kricka 1 – Nucleic Acid Hybridization Test Formats: Strategies and Applications , 1992 .

[8]  I. Willner,et al.  Probing Biomolecular Interactions at Conductive and Semiconductive Surfaces by Impedance Spectroscopy: Routes to Impedimetric Immunosensors, DNA‐Sensors, and Enzyme Biosensors , 2003 .

[9]  Tomoji Kawai,et al.  Immobilization of Probe DNA on Ta2O5 Thin Film and Detection of Hybridized Helix DNA using IS-FET , 2004 .

[10]  Andreas Offenhäusser,et al.  Possibilities and limitations of label-free detection of DNA hybridization with field-effect-based devices , 2005 .

[11]  F. Uslu,et al.  Labelfree fully electronic nucleic acid detection system based on a field-effect transistor device. , 2004, Biosensors & bioelectronics.

[12]  J. Watts,et al.  An Introduction to Surface Analysis by XPS and AES , 1990 .

[13]  Piet Bergveld,et al.  The future of biosensors , 1996 .

[14]  T. G. Drummond,et al.  Electrochemical DNA sensors , 2003, Nature Biotechnology.

[15]  G Zeck,et al.  Spatially resolved electronic detection of biopolymers. , 2004, Physical review. E, Statistical, nonlinear, and soft matter physics.

[16]  E. Souteyrand,et al.  DIRECT DETECTION OF THE HYBRIDIZATION OF SYNTHETIC HOMO-OLIGOMER DNA SEQUENCES BY FIELD EFFECT , 1997 .

[17]  Yuji Miyahara,et al.  DNA Analysis Chip Based on Field-Effect Transistors , 2005 .

[18]  Makoto Ishida,et al.  Fabrication and Characteristics of a Field Effect Transistor-Type Charge Sensor for Detecting Deoxyribonucleic Acid Sequence , 2003 .

[19]  Yuji Miyahara,et al.  Immobilization of oligonucleotide probes on Si3N4 surface and its application to genetic field effect transistor , 2004 .

[20]  S. Jamasb An analytical technique for counteracting drift in ion-selective field effect transistors (ISFETs) , 2004, IEEE Sensors Journal.

[21]  S. D. Collins,et al.  A physical model for drift in pH ISFETs , 1998 .

[22]  P. Bergveld,et al.  Operation of chemically sensitive field-effect sensors as a function of the insulator-electrolyte interface , 1983, IEEE Transactions on Electron Devices.

[23]  Jang-Kyoo Shin,et al.  An FET-type charge sensor for highly sensitive detection of DNA sequence. , 2004, Biosensors & bioelectronics.

[24]  P. Sorger,et al.  Electronic detection of DNA by its intrinsic molecular charge , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Joseph Wang Electrochemical nucleic acid biosensors , 2002 .

[26]  F. Pouthas,et al.  DNA detection on transistor arrays following mutation-specific enzymatic amplification , 2004 .

[27]  Y. Nemirovsky,et al.  Low frequency noise and drift in Ion Sensitive Field Effect Transistors , 2000 .