Detection of Human Interleukine‐2 Gene Using a Label‐Free Electrochemical DNA Hybridization Biosensor on the Basis of a Non‐Inosine Substituted Probe

The human interleukine-2 gene (hIL-2) is detected with a label-free DNA hybridization biosensor using a non-inosine substituted probe. The sensor relies on the immobilization of a 20-mer antisense single strand oligonucleotide (chIL-2) related to the human interleukine-2 gene on the pencil graphite electrode (PGE) as a probe. The guanine oxidation signal was monitored using anodic differential pulse voltammetry (ADPV). The electrochemical pretreatment of the polished PGE at 1.80 V for 5 min is suggested. Then, 5 min immobilization at 0.50 V was found as the optimum condition for immobilization of the probe. The electrochemical detection of hybridization between chIL-2 and hIL-2 as a target was accomplished. The selectivity of the biosensor was studied using noncomplementary oligonucleotides. Diagnostic performance of the biosensor is described and the detection limit is found 36 pg/mL.

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

[2]  A. Eckhardt,et al.  Probing biomolecule recognition with electron transfer: electrochemical sensors for DNA hybridization. , 1997, Bioconjugate chemistry.

[3]  Slobodan V. Jovanovic,et al.  How Easily Oxidizable Is DNA? One-Electron Reduction Potentials of Adenosine and Guanosine Radicals in Aqueous Solution , 1997 .

[4]  Kagan Kerman,et al.  Indicator‐Free Electrochemical DNA Biosensor Based on Adenine and Guanine Signals , 2002 .

[5]  G. Rivas,et al.  Adsorption and electrooxidation of nucleic acids at carbon nanotubes paste electrodes , 2004 .

[6]  M. Raffeld,et al.  Cancer Regression and Autoimmunity in Patients After Clonal Repopulation with Antitumor Lymphocytes , 2002, Science.

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

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

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

[10]  T. Merigan,et al.  Regulation by recombinant interleukin-2 of protective immunity against recurrent herpes simplex virus type 2 genital infection in guinea pigs , 1987, Journal of virology.

[11]  Shengshui Hu,et al.  Direct electrochemistry of DNA, guanine and adenine at a nanostructured film-modified electrode , 2003, Analytical and bioanalytical chemistry.

[12]  Anthony Guiseppi-Elie,et al.  Frequency dependent and surface characterization of DNA immobilization and hybridization. , 2004, Biosensors & bioelectronics.

[13]  Kagan Kerman,et al.  Detection of achondroplasia G380R mutation from PCR amplicons by using inosine modified carbon electrodes based on electrochemical DNA chip technology. , 2003, Clinica chimica acta; international journal of clinical chemistry.

[14]  Yuzuru Takamura,et al.  Peptide nucleic acid modified magnetic beads for intercalator based electrochemical detection of DNA hybridization , 2004 .

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

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

[17]  Yuzuru Takamura,et al.  Label-free electrochemical detection of DNA hybridization on gold electrode , 2003 .

[18]  R. Gallo,et al.  Selective in vitro growth of T lymphocytes from normal human bone marrows. , 1976, Science.

[19]  Anthony Guiseppi-Elie,et al.  Electrical frequency dependent characterization of DNA hybridization. , 2003, Biosensors & bioelectronics.

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

[21]  H. Thorp,et al.  Modification of indium tin oxide electrodes with repeat polynucleotides: electrochemical detection of trinucleotide repeat expansion. , 2001, Analytical chemistry.

[22]  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 .

[23]  J Wang,et al.  Magnetic bead-based label-free electrochemical detection of DNA hybridization. , 2001, The Analyst.

[24]  R. Engstrom.  Electrochemical pretreatment of glassy carbon electrodes , 1982 .

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

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

[27]  M. Hejazi,et al.  Developing an electrochemical deoxyribonucleic acid (DNA) biosensor on the basis of human interleukine-2 gene using an electroactive label. , 2006, Analytica chimica acta.

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

[29]  Giovanna Marrazza,et al.  Coupling of an indicator-free electrochemical DNA biosensor with polymerase chain reaction for the detection of DNA sequences related to the apolipoprotein E , 2002 .

[30]  E M Southern,et al.  Studies on the base pairing properties of deoxyinosine by solid phase hybridisation to oligonucleotides. , 1994, Nucleic acids research.

[31]  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.

[32]  Steven A. Rosenberg,et al.  Adoptive-cell-transfer therapy for the treatment of patients with cancer , 2003, Nature Reviews Cancer.

[33]  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.

[34]  A. Erdem,et al.  Rigid carbon composites: a new transducing material for label-free electrochemical genosensing , 2004 .