Rapid diagnostic barcode system for codetection of multiple protein markers

An ultrasensitive immunodiagnostic readout method based on an electrochemical analysis is presented. Different inorganic quantum dot (QD) nanocrystals (ZnS, CdS, and PbS) are tagged to antibodies for the on-site voltammetric stripping measurements of multiple antigen targets. The multiprotein electrical sensing capability is coupled to the amplification feature of anodic stripping voltammetric transduction and with an efficient magnetic removal (to minimize nonspecific adsorption and cross-reactivity effects). Sandwich-immunoassay formats were performed using model proteins (/spl beta//sub 2/-microglobulin, myoglobin, and human serum albumin). These encoding QD tracers with distinct redox potential yield highly sensitive and selective stripping peaks at -1.11 V (Zn), -0.67 V (Cd), and -0.52 V (Pb) at the mercury-film screen printed carbon electrode (versus Ag/AgCl reference). The position and size of these peaks reflect the identity and risk level of the corresponding antigen marker. The favorable signal-to-noise characteristics of the response for the initial 25-ng/mL mixture indicate a detection limit of ca. 10 ng/mL far below the early warning range and allow a reliable determination of very low protein concentrations. Such analog peaks of the QDs were converted to simple and rapid barcode signals. The digital readout system can code 215 electrically tuned barcodes to mark different protein analytes and to be useful for a wireless communication system.

[1]  B. Ahmed,et al.  Applications of barcode technology in automated storage and retrieval systems , 1996, Proceedings of the 1996 IEEE IECON. 22nd International Conference on Industrial Electronics, Control, and Instrumentation.

[2]  Igor L. Medintz,et al.  Multiplexed toxin analysis using four colors of quantum dot fluororeagents. , 2004, Analytical chemistry.

[3]  A. Casbard,et al.  Barcode technology: its role in increasing the safety of blood transfusion , 2003, Transfusion.

[4]  S. Nie,et al.  Quantum-dot-tagged microbeads for multiplexed optical coding of biomolecules , 2001, Nature Biotechnology.

[5]  W Richter,et al.  MR imaging contrast enhancement based on intermolecular zero quantum coherences. , 1998, Science.

[6]  Lu,et al.  Bismuth-coated carbon electrodes for anodic stripping voltammetry , 2000, Analytical chemistry.

[7]  J H Kim,et al.  Conductimetric membrane strip immunosensor with polyaniline-bound gold colloids as signal generator. , 2000, Biosensors & bioelectronics.

[8]  A. Klug,et al.  Alkyl Xanthates: New Capping Agents for Metal Colloids. Capping of Platinum Nanoparticles , 2001 .

[9]  Guodong Liu,et al.  Electrochemical coding technology for simultaneous detection of multiple DNA targets. , 2003, Journal of the American Chemical Society.

[10]  Shlomo Efrima,et al.  Synthesis of high-quality metal sulfide nanoparticles from alkyl xanthate single precursors in alkylamine solvents , 2003 .

[11]  Christine M. Micheel,et al.  Biological applications of colloidal nanocrystals , 2003 .

[12]  S. Nie,et al.  Molecular profiling of single cells and tissue specimens with quantum dots. , 2003, Trends in biotechnology.

[13]  S. Pathak,et al.  Hydroxylated quantum dots as luminescent probes for in situ hybridization. , 2001, Journal of the American Chemical Society.

[14]  T. Joos,et al.  Protein microarray technology. , 2002, Trends in biotechnology.

[15]  N. Pradhan,et al.  Single-precursor, one-pot versatile synthesis under near ambient conditions of tunable, single and dual band fluorescing metal sulfide nanoparticles. , 2003, Journal of the American Chemical Society.

[16]  H.M. Wechsler,et al.  Digital image processing, 2nd ed. , 1981, Proceedings of the IEEE.

[17]  S. Efrima,et al.  Xanthate Capping of Silver, Copper, and Gold Colloids , 2002 .

[18]  J. Church,et al.  The clinical application of a urine albumin:creatinine ratio point-of-care device. , 2001, Clinica chimica acta; international journal of clinical chemistry.

[19]  Joseph Wang,et al.  Electrochemical stripping detection of DNA hybridization based on cadmium sulfide nanoparticle tags , 2002 .

[20]  S. Nie,et al.  Quantum dot bioconjugates for ultrasensitive nonisotopic detection. , 1998, Science.

[21]  P. D'Orazio Biosensors in clinical chemistry. , 2003, Clinica chimica acta; international journal of clinical chemistry.

[22]  D. Balding,et al.  HLA Sequence Polymorphism and the Origin of Humans , 2006 .

[23]  K. Chia,et al.  Markers of diabetic nephropathy. , 1998, Journal of diabetes and its complications.

[24]  Jonathan Gressel,et al.  Universal inheritable barcodes for identifying organisms. , 2002, Trends in plant science.