Impedimetric detection of covalently attached biomolecules on field‐effect transistors

A method for impedimetric detection of biomolecules with field-effect transistor (FET) arrays is presented. For biomolecular detection, the corresponding molecules such as single-stranded DNA or bovine serum albumine (BSA) were covalently immobilized to the gate structure of 16-channel open-gate FET devices. A micro spotter system was used to site-selectively attach biomolecules to selected channels out of the array. The differential transistor transfer function (TTF) of the FETs with attached biomolecules compared to free FETs was recorded. With this impedimetric technique it was possible to reliably detect biomolecules, because the recordings were not disturbed by the typical long-term drift of the sensors like in potentiometric readout mode. For transistor gates, where DNA sequences of different length were attached, small differences in the TTF spectra were detected. When BSA was covalently immobilized to the FETs clear differences in the TTF spectra were detected, which were independent on buffer pH variations around the isoelectric point of the protein. Based on the results presented in this article it can be concluded, that the TTF method detects passive components of the biomolecules like resistance and capacitance rather than surface charge effects.

[1]  Constantin Polychronakos,et al.  Functionalization of Si/SiO2 substrates with homooligonucleotides for a DNA biosensor , 1999 .

[2]  J. Greve,et al.  Influence of an immunological precipitate on D.C. and A.C. behaviour of an isfet , 1989 .

[3]  B. Liedberg,et al.  A high-density poly(ethylene glycol) polymer brush for immobilization on glass-type surfaces. , 2000, Biosensors & bioelectronics.

[4]  S. Ingebrandt,et al.  Backside contacted field effect transistor array for extracellular signal recording. , 2003, Biosensors & bioelectronics.

[5]  S. Franzen,et al.  Probing BSA binding to citrate-coated gold nanoparticles and surfaces. , 2005, Langmuir : the ACS journal of surfaces and colloids.

[6]  R. Golbik,et al.  Protein adsorption and leakage in carrier–enzyme systems , 1991, Biotechnology and bioengineering.

[7]  I. Willner,et al.  Enzyme monolayer-functionalized field-effect transistors for biosensor applications , 2000 .

[8]  Charles M. Lieber,et al.  Nanowire-based biosensors. , 2006, Analytical chemistry.

[9]  Andreas Offenhäusser,et al.  Detection of DNA hybridization by a field‐effect transistor with covalently attached catcher molecules , 2006 .

[10]  S. Ingebrandt,et al.  Time-dependent observation of individual cellular binding events to field-effect transistors. , 2009, Biosensors & bioelectronics.

[11]  C. Consolandi,et al.  TWO EFFICIENT POLYMERIC CHEMICAL PLATFORMS FOR OLIGONUCLEOTIDE MICROARRAY PREPARATION , 2002, Nucleosides, nucleotides & nucleic acids.

[12]  P Bergveld,et al.  Development of an ion-sensitive solid-state device for neurophysiological measurements. , 1970, IEEE transactions on bio-medical engineering.

[13]  Andreas Offenhäusser,et al.  Surface activation of thin silicon oxides by wet cleaning and silanization , 2006 .

[14]  T. Bayer,et al.  Development of ion-sensitive field-effect transistor-based sensors for benzylphosphonic acids and thiophenols using molecularly imprinted TiO2 films , 2004 .

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

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

[17]  I. Willner,et al.  Imprinting of chiral molecular recognition sites in thin TiO2 films associated with field-effect transistors: novel functionalized devices for chiroselective and chirospecific analyses. , 2001, Chemistry.

[18]  A. van den Berg,et al.  Membrane characterization of anion-selective CHEMFETs by impedance spectroscopy. , 2000, Analytical chemistry.

[19]  Itamar Willner,et al.  Probing antigen-antibody binding processes by impedance measurements on ion-sensitive field-effect transistor devices and complementary surface plasmon resonance analyses: development of cholera toxin sensors. , 2002, Analytical chemistry.

[20]  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.

[21]  J. Feijen,et al.  Detection of charged proteins by means of impedance measurements , 1992 .

[22]  Michael J. Schöning,et al.  Label-free detection of charged macromolecules by using a field-effect-based sensor platform: Experiments and possible mechanisms of signal generation , 2007 .

[23]  I. Willner,et al.  Selective sensing of triazine herbicides in imprinted membranes using ion-sensitive field-effect transistors and microgravimetric quartz crystal microbalance measurements. , 2002, The Analyst.

[24]  Andreas Offenhäusser,et al.  Label‐free detection of DNA using field‐effect transistors , 2006 .

[25]  P Bergveld,et al.  A critical evaluation of direct electrical protein detection methods. , 1991, Biosensors & bioelectronics.

[26]  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 .

[27]  C. T. Aravindakumar,et al.  Transport studies of BSA, lysozyme and ovalbumin through chitosan/polystyrene sulfonate multilayer membrane , 2007 .

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

[29]  D. Reinhoudt,et al.  The effects of covalent binding of the electroactive components in durable CHEMFET membranes impedance spectroscopy and ion sensitivity studies , 1998 .

[30]  E Souteyrand,et al.  Optimisation of a silicon/silicon dioxide substrate for a fluorescence DNA microarray. , 2004, Biosensors & bioelectronics.

[31]  Itamar Willner,et al.  The Use of Impedance Spectroscopy for the Characterization of Protein-Modified ISFET Devices: Application of the Method for the Analysis of Biorecognition Processes , 2001 .

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

[33]  S. Ingebrandt,et al.  Label-free detection of single nucleotide polymorphisms utilizing the differential transfer function of field-effect transistors. , 2007, Biosensors & bioelectronics.

[34]  A. Dasgupta,et al.  Gold nanoparticle-based tool to study protein conformational variants: implications in hemoglobinopathy. , 2007, Nanomedicine : nanotechnology, biology, and medicine.

[35]  M. Schöning,et al.  Recent advances in biologically sensitive field-effect transistors (BioFETs). , 2002, The Analyst.

[36]  Piet Bergveld,et al.  Extracellular Potential Recordings by Means of a Field Effect Transistor Without Gate Metal, Called OSFET , 1976, IEEE Transactions on Biomedical Engineering.

[37]  I. Willner,et al.  Imprinting of nucleotide and monosaccharide recognition sites in acrylamidephenylboronic acid-acrylamide copolymer membranes associated with electronic transducers. , 2002, Analytical chemistry.