Detection of DNA and proteins using amorphous silicon ion-sensitive thin-film field effect transistors.

Amorphous silicon-based ion-sensitive field-effect transistors (a-Si:H ISFETs) are used for the label-free detection of biological molecules. The covalent immobilization of DNA, followed by DNA hybridization, and of the surface adsorption of oligonucleotides and proteins were detected electronically by the a-Si:H ISFET. The ISFET measurements are performed with an external Ag/AgCl microreference electrode immersed in 100mM phosphate buffer electrolyte with pH 7.0. Threshold voltage shifts in the transfer curve of the ISFETs are observed resulting from successive steps of surface chemical functionalization, covalent DNA attachment to the functionalized surface, surface blocking, and hybridization with a complementary target. The surface sensitivity achieved for DNA oligonucleotides is of the order of 1pmol/cm(2). Point-of-zero charge estimations were made for the functionalized surfaces and for the device surface after DNA immobilization and hybridization. The results show a correlation between the changes in the point-of-zero charge and the shift observed in the threshold voltage of the devices. Electronic detection of adsorbed proteins and DNA is also achieved by monitoring the shifts of the threshold voltage of the ISFETs, with a sensitivity of approximately 50nM.

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

[2]  V. Chu,et al.  Thin-film silicon MEMS DNA sensors , 2006 .

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

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

[5]  Y.-W. Kim,et al.  Statics and dynamics of strongly charged soft matter , 2005 .

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

[7]  B. Batlogg,et al.  Threshold voltage shift in organic field effect transistors by dipole monolayers on the gate insulator , 2004 .

[8]  D. Landheer,et al.  Calculation of the Response of Field-Effect Transistors to Charged Biological Molecules , 2007, IEEE Sensors Journal.

[9]  W. Birch,et al.  Molecular interactions between DNA and an aminated glass substrate. , 2003, Journal of colloid and interface science.

[10]  Hiroshi Iwasaki,et al.  Fabrication of Thin-Film LAPS with Amorphous Silicon , 2004, Sensors (Basel, Switzerland).

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

[12]  D. Richards,et al.  Labeling effects on the isoelectric point of green fluorescent protein. , 1999, Journal of chromatography. A.

[13]  R. Street,et al.  Hydrogenated amorphous silicon: Index , 1991 .

[14]  Label-free electronic detection of biomolecules using a-Si:H field-effect devices , 2006 .

[15]  George G Malliaras,et al.  Chemical and biological sensors based on organic thin-film transistors , 2005, Analytical and bioanalytical chemistry.

[16]  M. Stutzmann,et al.  pH sensors based on hydrogenated diamond surfaces , 2005 .

[17]  Domenico Caputo,et al.  Hydrogenated amorphous silicon ultraviolet sensor for deoxyribonucleic acid analysis , 2006 .

[18]  M. Stutzmann,et al.  Catalytic activity of enzymes immobilized on AlGaN/GaN solution gate field-effect transistors , 2006 .

[19]  Shunri Oda,et al.  Construction of amorphous silicon ISFET , 1989 .

[20]  Robert A. Street,et al.  Amorphous Silicon Electronics , 1992 .

[21]  V Chu,et al.  An on-chip thin film photodetector for the quantification of DNA probes and targets in microarrays. , 2004, Nucleic acids research.

[22]  Lester F. Eastman,et al.  pH response of GaN surfaces and its application for pH-sensitive field-effect transistors , 2003 .

[23]  K. Welinder Amino Acid Sequence Studies of Horseradish Peroxidase , 1979 .

[24]  H. Kawarada,et al.  Surface-modified Diamond Field-effect Transistors for Enzyme-immobilized Biosensors , 2004 .

[25]  Håkan Wennerström,et al.  The Colloidal Domain: Where Physics, Chemistry, Biology and Technology Meet , 1994 .

[26]  Eiichi Tamiya,et al.  Glucose sensor based on an amorphous silicon ISFET , 1989 .

[27]  H. Lüth,et al.  A (Bio-)Chemical Field-Effect Sensor with Macroporous Si as Substrate Material and a SiO2 / LPCVD-Si3N4 Double Layer as pH Transducer , 2002 .

[28]  R. Georgiadis,et al.  The effect of surface probe density on DNA hybridization. , 2001, Nucleic acids research.

[29]  Michael J. Schöning,et al.  Penicillin detection by means of field-effect based sensors: EnFET, capacitive EIS sensor or LAPS? , 2001 .

[30]  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.

[31]  G. Gonye,et al.  Characteristics of field-effect devices with gate oxide modification by DNA , 2005 .

[32]  G. S. Manning Counterion condensation on a helical charge lattice , 2001 .