Developing an electrochemical deoxyribonucleic acid (DNA) biosensor on the basis of human interleukine-2 gene using an electroactive label.

Development of an electrochemical DNA biosensor based on a human interleukine-2 (IL-2) gene probe, using a pencil graphite electrode (PGE) as transducer and methylene blue (MB) as electroactive label is described. The sensor relies on the immobilization of a 20-mer single stranded oligonucleotide probe (hIL-2) related to the IL-2 gene on the electrode. The hybridization between the probe and its complementary sequence (chIL-2) as the target was studied by square wave voltammetry (SWV) of MB accumulated on the PGE. In this approach the extent of hybridization is evaluated on the basis of the difference between SWV signals of MB accumulated on the probe-PGE and MB accumulated on the probe-target-PGE. Some hybridization experiments with non-complementary oligonucleotides were carried out to assess whether the suggested DNA sensor responds selectively to the target. Some experimental variables affecting the performance of the biosensor including: polishing of PGE, its electrochemical activation conditions (i.e., activation potential and activation time) and probe immobilization conditions on the electrodes (i.e., immobilization potential and time) were investigated and the optimum values of 1.80 V and 300 s for PGE activation, and -0.5 V and 400s for the probe immobilization on the electrode were suggested.

[1]  A. Merkoçi,et al.  Electrochemical genosensor design: immobilisation of oligonucleotides onto transducer surfaces and detection methods. , 2000, Biosensors & bioelectronics.

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

[3]  K. M. Millan,et al.  Voltammetric DNA biosensor for cystic fibrosis based on a modified carbon paste electrode. , 1994, Analytical chemistry.

[4]  Gustavo Rivas,et al.  Indicator-free electrochemical DNA hybridization biosensor , 1998 .

[5]  Kagan Kerman,et al.  Allele-specific genotype detection of factor V Leiden mutation from polymerase chain reaction amplicons based on label-free electrochemical genosensor. , 2002, Analytical chemistry.

[6]  W. Fiers,et al.  Production of soluble and active recombinant murine interleukin-2 in Escherichia coli: high level expression, Kil-induced release, and purification. , 1995, Protein expression and purification.

[7]  Mehmet Ozsoz,et al.  Electrochemical DNA biosensor for the detection and discrimination of herpes simplex Type I and Type II viruses from PCR amplified real samples , 2004 .

[8]  Qun Zhou,et al.  Viral vectors for cancer gene therapy: viral dissemination and tumor targeting. , 2005, Current gene therapy.

[9]  K. M. Millan,et al.  Sequence-selective biosensor for DNA based on electroactive hybridization indicators. , 1993, Analytical chemistry.

[10]  宁北芳,et al.  疟原虫var基因转换速率变化导致抗原变异[英]／Paul H, Robert P, Christodoulou Z, et al//Proc Natl Acad Sci U S A , 2005 .

[11]  Joseph Wang,et al.  Carbon-nanotube-modified glassy carbon electrodes for amplified label-free electrochemical detection of DNA hybridization. , 2003, The Analyst.

[12]  Kagan Kerman,et al.  Electrochemical DNA biosensor for the detection of specific gene related to Microcystis species , 2001 .

[13]  Adam Heller,et al.  Screen printing of nucleic acid detecting carbon electrodes. , 2002, Analytical chemistry.

[14]  B. Hames,et al.  Nucleic acid hybridisation : a practical approach , 1985 .

[15]  D. Cerretti,et al.  Cloning, sequence, and expression of bovine interleukin 2. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[16]  D. Pang,et al.  Micromethod for the investigation of the interactions between DNA and redox-active molecules. , 1998, Analytical chemistry.

[17]  Ningning Zhu,et al.  Electrochemical detection of DNA hybridization using methylene blue and electro-deposited zirconia thin films on gold electrodes , 2004 .

[18]  K. Kerman,et al.  Detection of Interaction Between Metal Complex Indicator and DNA by Using Electrochemical Biosensor , 1999 .

[19]  Mehmet Ozsoz,et al.  Disposable electrochemical biosensor for the detection of the interaction between DNA and lycorine based on guanine and adenine signals. , 2003, Journal of pharmaceutical and biomedical analysis.

[20]  Jan Tavernier,et al.  Molecular cloning of human interleukin 2 cDNA and its expression in E. coli , 1983, Nucleic Acids Res..

[21]  E. Paleček,et al.  Electrochemistry of Nucleic Acids and Development of DNA Sensors , 2002 .

[22]  Kagan Kerman,et al.  Novel hybridization indicator methylene blue for the electrochemical detection of short DNA sequences related to the hepatitis B virus , 2000 .

[23]  J. Rothel,et al.  Expression of ovine interleukin-2 cDNA in Escherichia coli. , 1997, Veterinary immunology and immunopathology.

[24]  J. Justin Gooding,et al.  Voltammetric determination of DNA hybridization using methylene blue and self-assembled alkanethiol monolayer on gold electrodes , 2002 .

[25]  Masato Saito,et al.  Electrochemical DNA quantification based on aggregation induced by Hoechst 33258 , 2004 .

[26]  Rene Kizek,et al.  Application of avidin-biotin technology and adsorptive transfer stripping square-wave voltammetry for detection of DNA hybridization and avidin in transgenic avidin maize. , 2003, Analytical chemistry.

[27]  Kagan Kerman,et al.  Methylene Blue as a Novel Electrochemical Hybridization Indicator , 2001 .

[28]  Yan Lin,et al.  Expression of an interleukin-6 - interleukin-2 fusion protein (pIL-6-IL-2) in P. pastoris. , 2004, European cytokine network.

[29]  P. A. Ropp,et al.  Toward electrochemical resolution of two genes on one electrode: using 7-deaza analogues of guanine and adenine to prepare PCR products with differential redox activity. , 2002, Analytical chemistry.

[30]  L T Kubota,et al.  Polishable and renewable DNA hybridization biosensors. , 1998, Analytical chemistry.

[31]  Elena Komarova,et al.  Direct electrochemical sensor for fast reagent-free DNA detection. , 2005, Biosensors & bioelectronics.

[32]  E. Paleček,et al.  Trace Measurements of RNA by Potentiometric Stripping Analysis at Carbon Paste Electrodes , 1995 .

[33]  Marco Mascini,et al.  Electrochemical DNA biosensor for the detection of TT and Hepatitis B virus from PCR amplified real samples by using methylene blue. , 2002, Talanta.

[34]  A. Erdem,et al.  Interaction of the anticancer drug epirubicin with DNA , 2001 .

[35]  A J Thomson,et al.  Methylene blue as an electrochemical discriminator of single- and double-stranded oligonucleotides immobilised on gold substrates. , 2001, The Analyst.

[36]  J. Justin Gooding,et al.  Evidence for the direct interaction between methylene blue and guanine bases using DNA-modified carbon paste electrodes , 2002 .

[37]  Christian Soeller,et al.  Label-free electrochemical DNA sensor based on functionalised conducting copolymer. , 2005, Biosensors & bioelectronics.

[38]  R. Robb.  Interleukin 2: the molecule and its function. , 1984, Immunology today.

[39]  H. Katinger,et al.  Generation of an Influenza A Virus Vector Expressing Biologically Active Human Interleukin-2 from the NS Gene Segment , 2005, Journal of Virology.

[40]  Kagan Kerman,et al.  Electrochemical genosensor for the detection of interaction between methylene blue and DNA , 2002 .

[41]  Xianting Zeng,et al.  Electrochemical biosensing of DNA with capture probe covalently immobilized onto glassy carbon surface , 2005 .

[42]  Sister Mary Berenice.  Facts and Comparisons , 1951 .

[43]  W. Heineman,et al.  Laboratory techniques in electroanalytical chemistry , 1984 .

[44]  J. Justin Gooding,et al.  Electrochemical detection of hybridization using peptide nucleic acids and methylene blue on self-assembled alkanethiol monolayer modified gold electrodes , 2002 .

[45]  E. Paleček,et al.  Interactions of antitumor drug daunomycin with DNA in solution and at the surface , 1998 .

[46]  F. Jelen,et al.  DNA and PNA sensing on mercury and carbon electrodes by using methylene blue as an electrochemical label. , 2002, Bioelectrochemistry.

[47]  Joseph Wang,et al.  Pencil-based renewable biosensor for label-free electrochemical detection of DNA hybridization , 2001 .

[48]  Elizabeth M. Boon,et al.  Single-base mismatch detection based on charge transduction through DNA. , 1999, Nucleic acids research.

[49]  S. Paul,et al.  Gene-based vaccines and immunotherapeutics , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[50]  J Wang,et al.  Renewable pencil electrodes for highly sensitive stripping potentiometric measurements of DNA and RNA. , 2000, The Analyst.

[51]  J Wang,et al.  DNA electrochemical biosensor for the detection of short DNA sequences related to the human immunodeficiency virus. , 1996, Analytical chemistry.

[52]  E. Paleček,et al.  Cyclic voltammetry of echinomycin and its interaction with double-stranded and single-stranded DNA adsorbed at the electrode. , 2002, Bioelectrochemistry.

[53]  G. Marrazza,et al.  Carbon and gold electrodes as electrochemical transducers for DNA hybridisation sensors. , 2004, Biosensors & bioelectronics.

[54]  D. Regier,et al.  Design, synthesis and expression of a human interleukin-2 gene incorporating the codon usage bias found in highly expressed Escherichia coli genes. , 1988, Nucleic acids research.

[55]  H. Ju,et al.  Interaction between nile blue and immobilized single- or double-stranded DNA and its application in electrochemical recognition , 2005 .

[56]  Erwin K. Kastrup,et al.  Drug facts and comparisons , 1977 .

[57]  Eric P. Achterberg,et al.  Laboratory techniques in electroanalytical chemistry , 1996 .

[58]  M. Dequaire,et al.  Covalent immobilization of oligonucleotides on p-aminophenyl-modified carbon screen-printed electrodes for viral DNA sensing. , 2003, Chemical communications.