An amperometric chloramphenicol immunosensor based on cadmium sulfide nanoparticles modified-dendrimer bonded conducting polymer.

An amperometric chloramphenicol (CAP) immunosensor was fabricated by covalently immobilizing anti-chloramphenicol acetyl transferase (anti-CAT) antibody on cadmium sulfide nanoparticles (CdS) modified-dendrimer that was bonded to the conducting polymer (poly 5, 2': 5', 2''-terthiophene-3'-carboxyl acid (poly-TTCA)) layer. The AuNPs, dendrimers, and CdS nanoparticles were deposited onto the polymer layer in order to enhance the sensitivity of the sensor probes. The particle sizes were determined using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The immobilization of dendrimers, CdS, and anti-CAT were confirmed using energy disruptive spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), and quartz crystal microbalance (QCM) techniques. The detection of CAP was based on the competitive immuno-interaction between the free- and labeled-CAP for active sites of the anti-CAT. Hydrazine was used as the label for CAP, and it electrochemically catalyzed the reduction of H(2)O(2) at -0.35 V vs. Ag/AgCl. Under optimized conditions, the proposed immunosensor exhibited a linear range of CAP detection between 50 pg/mL and 950 pg/mL, and the detection limit was 45 pg/mL. The immunosensor was examined in real meat samples for the analysis of CAP.

[1]  D. Tomalia Birth of a new macromolecular architecture: dendrimers as quantized building blocks for nanoscale synthetic polymer chemistry , 2005 .

[2]  Yoon-Bo Shim,et al.  Disposable amperometric immunosensor system for rabbit IgG using a conducting polymer modified screen-printed electrode. , 2003, Biosensors & bioelectronics.

[3]  Shi-quan Han,et al.  Chemiluminescence immunoassay for chloramphenicol , 2005, Analytical and bioanalytical chemistry.

[4]  T. Izard,et al.  Structural basis for chloramphenicol tolerance in Streptomyces venezuelae by chloramphenicol phosphotransferase activity , 2001, Protein science : a publication of the Protein Society.

[5]  Joseph Wang Nanomaterial-based amplified transduction of biomolecular interactions. , 2005, Small.

[6]  J. Louvois Factors influencing the assay of antimicrobial drugs in clinical samples by the agar plate diffusion method , 1982 .

[7]  Muhammad J A Shiddiky,et al.  Hydrazine-catalyzed ultrasensitive detection of DNA and proteins. , 2007, Analytical chemistry.

[8]  A. Di Corcia,et al.  Liquid chromatographic-mass spectrometric methods for analyzing antibiotic and antibacterial agents in animal food products. , 2002, Journal of chromatography. A.

[9]  A. Alivisatos,et al.  Hybrid Nanorod-Polymer Solar Cells , 2002, Science.

[10]  R. M. Smith,et al.  Chloromycetin, a New Antibiotic From a Soil Actinomycete. , 1947, Science.

[11]  Interactions between dendrimers and charged probe molecules. 1. Theoretical methods for simulating proton and metal ion binding to symmetric polydentate ligands , 2002 .

[12]  Guodong Liu,et al.  Sensitive immunoassay of a biomarker tumor necrosis factor-alpha based on poly(guanine)-functionalized silica nanoparticle label. , 2006, Analytical chemistry.

[13]  Thomas J. White,et al.  Chloramphenicol: an Enzymological Microassay , 1976, Antimicrobial Agents and Chemotherapy.

[14]  J. Matthew Mauro,et al.  Self-Assembly of CdSe−ZnS Quantum Dot Bioconjugates Using an Engineered Recombinant Protein , 2000 .

[15]  Jun‐Jie Zhu,et al.  Electrochemiluminescence of CdSe quantum dots for immunosensing of human prealbumin. , 2008, Biosensors & bioelectronics.

[16]  William R. Heineman,et al.  A nanotube array immunosensor for direct electrochemical detection of antigen–antibody binding , 2007 .

[17]  B. Limoges,et al.  An electrochemical metalloimmunoassay based on a colloidal gold label. , 2000, Analytical chemistry.

[18]  Y. Shim,et al.  Direct DNA hybridization detection based on the oligonucleotide-functionalized conductive polymer. , 2001, Analytical chemistry.

[19]  I. Park,et al.  Development of a direct-binding chloramphenicol sensor based on thiol or sulfide mediated self-assembled antibody monolayers. , 2004, Biosensors & bioelectronics.

[20]  P. Ugo,et al.  Nanoelectrode ensembles as recognition platform for electrochemical immunosensors. , 2008, Biosensors & bioelectronics.

[21]  T. Ohsaka,et al.  An extraordinary electrocatalytic reduction of oxygen on gold nanoparticles-electrodeposited gold electrodes ☆ , 2002 .

[22]  Namsoo Kim,et al.  Development of a chemiluminescent immunosensor for chloramphenicol. , 2006, Analytica chimica acta.

[23]  M. Borkovec,et al.  Microscopic protonation equilibria of Poly(amidoamine) dendrimers from macroscopic titrations , 2003 .

[24]  Flora Boccuzzi,et al.  Au/TiO2 nanostructured catalyst: effects of gold particle sizes on CO oxidation at 90 K , 2001 .

[25]  Guodong Liu,et al.  Electrochemical quantification of single-nucleotide polymorphisms using nanoparticle probes. , 2007, Journal of the American Chemical Society.

[26]  Y. Shim,et al.  An amperometric immunosensor for osteoproteogerin based on gold nanoparticles deposited conducting polymer. , 2008, Biosensors & bioelectronics.

[27]  Y. Shim,et al.  Simple preparation of terthiophene-3′-carboxylic acid and characterization of its polymer , 2002 .

[28]  Y. Shim,et al.  The potential use of hydrazine as an alternative to peroxidase in a biosensor: comparison between hydrazine and HRP-based glucose sensors. , 2005, Biosensors & bioelectronics.

[29]  D. Berry Reversed-phase high-performance liquid chromatographic determination of chloramphenicol in small biological samples. , 1987, Journal of chromatography.

[30]  Yun Xiang,et al.  Quantum-dot/aptamer-based ultrasensitive multi-analyte electrochemical biosensor. , 2006, Journal of the American Chemical Society.