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protein microarrays with carbon nanotubes as multicolor Raman labels

NATURE BIOTECHNOLOGY 2008年第11期26卷 1285-1292页

作者： Chen, Zhuo Tabakman, Scott M. Goodwin, Andrew P. Kattah, Michael G. Daranciang, Dan Wang, Xinran Zhang, Guangyu Li, Xiaolin Liu, Zhuang Utz, Paul J. Jiang, Kaili Fan, Shoushan Dai, Hongjie Stanford Univ Dept Chem Stanford CA 94305 USA Stanford Univ Adv Mat Lab Stanford CA 94305 USA Stanford Univ Sch Med Stanford CA 94305 USA Tsinghua Univ Dept Phys Tsinghua Foxconn Nanotechnol Res Ctr Beijing 100084 Peoples R China

The current sensitivity of standard fluorescence-based protein detection limits the use of protein arrays in research and clinical diagnosis. Here, we use functionalized, macromolecular single-walled carbon nanotubes ( SWNTs) as multicolor Raman labels for highly sensitive, multiplexed protein detection in an arrayed format. Unlike fluorescence methods, Raman detection benefits from the sharp scattering peaks of SWNTs with minimal background interference, affording a high signal-to-noise ratio needed for ultra-sensitive detection. When combined with surface-enhanced Raman scattering substrates, the strong Raman intensity of SWNT tags affords protein detection sensitivity in sandwich assays down to 1 fM-a three-order-of-magnitude improvement over most reports of fluorescence-based detection. We use SWNT Raman tags to detect human autoantibodies against proteinase 3, a biomarker for the autoimmune disease Wegener's granulomatosis, diluted up to 10(7)-fold in 1% human serum. SWNT Raman tags are not subject to photobleaching or quenching. By conjugating different antibodies to pure C-12 and C-13 SWNT isotopes, we demonstrate multiplexed two-color SWNT Raman- based protein detection.

关键词： protein microarrays CARBON nanotubes RAMAN spectroscopy FLUORESCENCE SUBSTRATES (Materials science) IMMUNOGLOBULINS RESEARCH BIOCHEMICAL markers WEGENER'S granulomatosis

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protein microarrays: new tools for pharmaceutical development

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ANALYTICAL AND BIOANALYTICAL CHEMISTRY 2003年第5期377卷 812-819页

作者： Kumble, KD Pointilliste Inc Mountain View CA 94043 USA

protein microarrays are a relatively new technology, which will dramatically impact the pharmaceutical industry. The critical need for more rapid identification of novel drug targets, and for obtaining high-quality information early in the target validation process is a major driver for the industry. High-throughput protein analytical techniques are critical for obtaining biological information beyond that which transcript analysis can provide, given that proteins are the "worker bees" in cells. The vast complexity of proteins when compared to DNA and RNA in terms of sheer number, and structural and biochemical diversity requires a higher degree of sophistication in both assay design and data analysis. High-throughput microarray technology platforms allow for simultaneous, multi-parametric analysis of complex protein mixtures. protein microarrays have tremendous potential as a tool for the study of protein - protein, enzyme - substrate, and antibody - antigen interactions among others. They can also be used for biomarkers and drug target identification via comparative proteomic analysis of healthy and disease tissues. More recently, cellular microarrays that enable identification of cell-surface receptors and other cell-surface proteins allowing rapid screening of cell-specific, novel drug targets, are being developed. This review will focus on the technical issues and potential applications of protein microarrays in pharmaceutical discovery.

关键词： protein microarrays IMMOBILIZED proteins proteinS DRUGS PHARMACEUTICAL industry NECTARIVORES

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protein microarrays and quantum dot probes for early cancer detection

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COLLOIDS AND SURFACES B-BIOINTERFACES 2007年第2期58卷 309-314页

作者： Zajac, Aleksandra Song, Dansheng Qian, Wei Zhukov, Tatyana Univ S Florida H Lee Moffitt Canc Ctr Dept Interdisciplinary Oncol Div Canc Prevent & Control Tampa FL 33612 USA Univ S Florida Res Inst Tampa FL 33612 USA

We describe here a novel approach for detection of cancer markers using quantum dot protein microarrays. Both relatively new technologies;quantum dots and protein microarrays, offer very unique features that together allow detection of cancer markers in biological specimens (serum, plasma, body fluids) at pg/ml concentration. Quantum dots offer remarkable photostability and brightness. They do not exhibit photobleaching common to organic fluorophores. Moreover, the high emission amplitude for QDs results in a marked improvement in the signal to noise ratio of the final image. protein microarrays allow highly parallel quantitation of specific proteins in a rapid, low-cost and low sample volume format. Furthermore the multiplexed assay enables detection of many proteins at once in one sample, making it a powerful tool for biomarker analysis and early cancer diagnostics. In a series of multiplexing experiments we investigated ability of the platform to detect six different cytokines in protein solution. We were able to detect TNF-alpha. IL-8, IL-6, MIP-1 beta, IL-13 and IL-1 beta down to picomolar concentration, demonstrating high sensitivity of the investigated detection system. We have also constructed and investigated two different models of quantum dot probes. One by conjugation of nanocrystals to antibody specific to the selected marker-IL-10, and the second by use of streptavidin coated quantum dots and biotinylated detector antibody. Comparison of those two models showed better performance of streptavidin QD-biotinylated detector antibody model. Data quantitated using custom designed computer program (CDAS) show that proposed methodology allows monitoring of changes in biomarker concentration in physiological range. (C) 2007 Elsevier B.V. All rights reserved.

关键词： quantum dots protein microarrays biomarkers cytokines cancer detection

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protein microarrays for diagnostic assays

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ANALYTICAL AND BIOANALYTICAL CHEMISTRY 2009年第5期393卷 1407-1416页

作者： Hartmann, Michael Roeraade, Johan Stoll, Dieter Templin, Markus F. Joos, Thomas O. Univ Tubingen NMI Nat & Med Sci Inst D-72770 Reutlingen Germany Royal Inst Technol Sch Chem Sci & Engn Dept Analyt Chem S-10044 Stockholm Sweden

protein microarray technology has enormous potential for in vitro diagnostics (IVD). Miniaturized parallelized immunoassays are perfectly suited to generating a maximum of diagnostically relevant information from minute amounts of sample whilst only requiring small amounts of reagent. protein microarrays have become well-established research tools in basic and applied research and the first products are already on the market. This article reviews the current state of protein microarrays and discusses developments and future demands relating to protein arrays in their role as multiplexed immunoassays in the field of diagnostics.

关键词： protein microarrays Multiplexed diagnostics In-vitro diagnostics Focused protein-profiling Analytical microarrays

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protein microarrays on carboxymethylated dextran hydrogels: Immobilization, characterization and application

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MICROCHIMICA ACTA 2004年第1-2期147卷 21-30页

作者： Zhou, Y Andersson, O Lindberg, P Liedberg, B Linkoping Univ S SENCE S-58183 Linkoping Sweden Linkoping Univ Div Sensor Sci S-58183 Linkoping Sweden Biacore AB S-75450 Uppsala Sweden

Tetraoctadecylammonium bromide (TOAB, (CH3(CH2)(17))(4)N+Br-) has been used to print temporary hydrophobic barriers on carboxymethylated dextran (CMD) hydrogels to create a generic platform for protein microarray applications. The primary reason for printing temporary hydrophobic barriers is to prevent cross-contamination and overflow during microdrop dispensing. Equally important is to eliminate the risk for non-specific binding to the barriers during analyte exposure. This has been accomplished by introducing a regeneration step that removes the barriers after ligand immobilization. The overall fabrication process was characterized by microscopic wetting, atomic force microscopy, imaging ellipsometry, fluorescence microscopy, surface plasmon microscopy and biospecific interaction analysis. A series of model proteins including transferrin, protein A, anti-myoglobin and bovine serum albumin was spotted into the TOAB-defined areas under different experimental conditions, e.g. at increased humidity and reduced substrate temperature or with glycerol as an additive in the protein solution. Much emphasis was devoted to studies aiming at exploring the homogeneity and activity of the immobilized proteins. The printed barriers were removed after protein immobilization using tert-n-butyl alcohol (TBA). TBA was found to be a very efficient agent as compared to previously used salt regeneration solutions, and the regeneration time could be reduced from similar to30 to 10 minutes. Finally, the potential of using the well established CMD hydrogel chemistry as a platform for protein microarrays was exploited using surface plasmon microscopy.

关键词： reversible hydrophobic barrier microcontact printing piezodispensing protein microarrays surface plasmon microscopy

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protein microarrays ON MIDTRIMESTER AMNIOTIC FLUIDS: A NOVEL APPROACH FOR THE DIAGNOSIS OF EARLY INTRAUTERINE INFLAMMATION RELATED TO PRETERM DELIVERY

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INTERNATIONAL JOURNAL OF IMMUNOPATHOLOGY AND PHARMACOLOGY 2012年第4期25卷 1029-1040页

作者： La Sala, G. B. Ardizzoni, A. Capodanno, F. Manca, L. Baschieri, M. C. Soncini, E. Peppoloni, S. Blasi, E. Arcispedale S Maria Nuova Ist Tecnol Avanzate & Modelli Assistenziali Oncol Dept Obstet & Gynecol Reggio Emilia Italy Univ Modena & Reggio Emilia Dept Diagnost Clin & Publ Hlth Med Ist Biol I-41125 Modena Italy

Novel technologies that allow simultaneous assessment of multiple biomarkers provide new and promising diagnostic/prognostic approaches. By protein microarrays, here we analyzed amniotic fluids (AF) from 50 women with preterm delivery (PTD) and 50 control women, who delivered at term. In detail, cytokines, chemokines, matrix metalloproteinases and antigen-specific antibodies were assessed. The AF analysis showed significant differences between women with preterm and term delivery in the levels of IL-1 alpha, IL-1 beta, IL-4, IL-6, IL-8, MCP-1, IFN-gamma and anti-HSV2 IgG. No significant differences were observed in the levels of TNF-alpha, MMP-2, MMP-9 and specific IgG for seven vertically transmitted pathogens. In conclusion, we demonstrated the feasibility of protein microarrays in the diagnosis of early intrauterine inflammation. The significant association between the increased levels of certain cytokines and preterm delivery argues on their relevance as early pathogenetic markers for identification of risk patients.

关键词： intrauterine inflammation protein microarrays term delivery preterm delivery amniotic fluid markers

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protein microarrays: Meeting analytical challenges for clinical applications

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CANCER CELL 2003年第4期3卷 317-325页

作者： Liotta, LA Espina, V Mehta, AI Calvert, V Rosenblatt, K Geho, D Munson, PJ Young, L Wulfkuhle, J Petricoin, EF NCI FDA NCI Clin Proteom Program Pathol Lab Canc Res Ctr Bethesda MD 20892 USA NCI FDA NCI Clin Proteom Program Ctr Biol Evaluat & Res Bethesda MD 20892 USA NIH Math & Stat Comp Lab Ctr Informat Technol Bethesda MD 20892 USA Howard Hughes Med Inst Bethesda MD 20892 USA

protein microarrays, one emerging class of proteomic technologies, have broad applications for discovery and quantitative analysis. A rapidly expanding use of this technology is the acquisition of information about the posttranslational modifications of proteins reflecting the activity state of signal pathways and networks, and is now employed for the analysis of biopsy samples in clinical trial research.

关键词： protein measurement Clinical Analysis of substances Employed protein microarrays EMPLOYMENT Biopsy sample Medical Clinical applications Post-Translational protein Processing Signal Pathways activity state Clinical Trial

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protein microarrays as tools for functional proteomics

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CURRENT OPINION IN CHEMICAL BIOLOGY 2005年第1期9卷 14-19页

作者： LaBaer, J Ramachandran, N Harvard Univ Sch Med Harvard Inst Proteom Dept Biol Chem & Mol Pharmacol Cambridge MA 02141 USA

protein microarrays present an innovative and versatile approach to study protein abundance and function at an unprecedented scale. Given the chemical and structural complexity of the proteome, the development of protein microarrays has been challenging. Despite these challenges there has been a marked increase in the use of protein microarrays to map interactions of proteins with various other molecules, and to identify potential disease biomarkers, especially in the area of cancer biology. In this review, we discuss some of the promising advances made in the development and use of protein microarrays.

关键词： protein microarrays PROTEOMICS proteinS MOLECULES BIOCHEMICAL markers

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protein microarrays: challenges and promises

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PHARMACOGENOMICS 2002年第4期3卷 527-536页

作者： Talapatra, A Rouse, R Hardiman, G Univ Calif San Diego Div Biol Biomed Genom Microarray Facil La Jolla CA 92093 USA Scripps Res Inst Dept Vasc Biol La Jolla CA 92037 USA

Genomics and proteomics are playing increasingly important roles as discovery tools in basic biological sciences and as diagnostic and rational therapeutic aids in the clinical arena. In recent years, high-density arrays of specified DNA sequences have gained popularity. protein microarrays are at the forefront of this biochip revolution and promise the parallel examination of large numbers of proteins. These miniaturized arrays are currently being developed to facilitate high analytical resolution, detection sensitivity and sample throughput. Many challenges are presented by proteome scale manipulation of proteins, as there is currently no methodological equivalent to the gene chip for comparative proteomics.

关键词： antibodies comparative proteomics expression analyses protein microarrays proteome

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protein microarrays to study carbohydrate-recognition events

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BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 2006年第19期16卷 5132-5135页

作者： Lee, Myung-ryul Park, Sungjin Shin, Injae Yonsei Univ Dept Chem Seoul 120749 South Korea

In order to expand areas in which protein microarrays can be used to solve important biological problems, we have investigated ways in which the technique can be employed for functional glycomics. Initially, our protein microarrays were used for the rapid identification of carbohydrate-binding proteins using trifunctional carbohydrate probes and fluorescent dye-labeled polysaccharides. Glycan probes were selectively bound to the corresponding lectins immobilized on the solid surface. In addition, these microarrays were also employed for profiling of carbohydrates on Jurkat T-cell surfaces. These cells adhered to ConA, RCA(120), SNA and WGA, indicating expression of alpha-Man, Gal, NeuNAc alpha 2,6Gal and GlcNAc residues on their surfaces. Furthermore, we determined binding affinities between WGA and carbohydrates by measuring IC50 values of GlcNAc that inhibited 50% of trivalent GlcNAc binding to WGA immobilized on the solid surface. All the experiments show that protein microarrays can be used to study carbohydrate-recognition events in the field of glycomics. (c) 2006 Elsevier Ltd. All rights reserved.

关键词： carbohydrates carbohydrate-protein interactions immobilization lectins protein microarrays

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