Protein microarrays and proteomics

MacBeath, Gavin

doi:10.1038/ng1037

Cited by 790 publications

(515 citation statements)

References 47 publications

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“…Each agent captures its target protein from a complex mixture (such as serum or a cell lysate), and the captured proteins are subsequently detected and quantified. 8 Although originally fluorescent dyes were applied for differential display of binding to a protein array, advancing MS technologies have allowed new approaches. One of these is surface enhanced laser desorption/ionization (SELDI)-TOF MS, 9 which is often Figure 1 Representative proteomic strategies.…”

Section: Proteome Analysismentioning

confidence: 99%

Proteome analysis of prostate cancer

Kuruma

Egawa

Ohishi

et al. 2004

Prostate Cancer Prostatic Dis

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In this paper, we briefly review cancer proteomics in general, with particular attention to our proteome analyses of prostate cancer. Our efforts include development of new tools and novel approaches to discovering proteins potentially useful as cancer diagnostic and/or prognostic biomarkers or as therapeutic targets. To this end, we analyzed prostate cancer proteomes using twodimensional gel electrophoresis employing agarose gels for the initial isoelectric focusing step (agarose 2-DE), with mass spectrometry used for protein identification. Agarose 2-DE offers advantages over the more widely used immobilized pH gradient 2-DE for separating high molecular mass proteins (15-500 kDa), thereby increasing its power to detect changes in the cancer's high-molecular mass proteomes.

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Section: Proteome Analysismentioning

confidence: 99%

Proteome analysis of prostate cancer

Kuruma

Egawa

Ohishi

et al. 2004

Prostate Cancer Prostatic Dis

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“…Numerous proteomic methods have been developed in the past twenty years to aid us in our understanding of enzyme function in biological processes and human disease states, including cancer [7][8][9]. However, many of these techniques, such as two-dimensional gel electrophoresis [8] or isotope-coded affinity-tagging [7], only focus on measuring changes in protein abundance.…”

Section: Introductionmentioning

confidence: 99%

“…Since most enzymes are expressed as inactive zymogens or reside in complex with their endogenous inhibitors, protein abundance does not necessarily correlate with activity. Other methods, such as protein microarrays [9], can provide information about an enzyme's activity state but generally require recombinantly expressed proteins that are monitored in isolation. Therefore, these technologies do not provide us with a functional understanding of native proteins in the physiologically relevant environment of a cell or whole organism.…”

Section: Introductionmentioning

confidence: 99%

Application of activity-based probes to the study of enzymes involved in cancer progression

Paulick

Bogyo

2008

Current Opinion in Genetics & Development

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SummaryMany tumor cells have elevated levels of hydrolytic and proteolytic enzymes, presumably to aid in key processes such as angiogenesis, cancer cell invasion, and metastasis. Since the activities of these enzymes are often regulated by post-translational mechanisms, their functional roles in cancer progression are difficult to study using traditional genomic and proteomic methods. Thus, methods that allow for the direct monitoring of enzyme activity in a physiologically relevant environment are required to better understand the roles of specific players in the complex process of tumorigenesis. This review highlights advances in the field of activity-based proteomics, which uses small molecules known as activity-based probes (ABPs) that covalently bind to the catalytic site of target enzymes. We discuss the application of ABPs to cancer biology, specifically to the discovery of tumor biomarkers, the screening of enzyme inhibitors, and the imaging of enzymes implicated in cancer.

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“…One of such analyses is the high-throughput measurement allowed by patterning the protein in a microarray format [1][2][3][4][5]. The proteins can be covalently linked to or immobilized by highcapacity absorption on a substrate surface, then detected with immunochemistry.…”

Section: Introductionmentioning

confidence: 99%

“…The most popular detection method uses fluorescence detection by specifically labeling the adsorbed proteins [4]. Conventional dyes currently present several shortcomings.…”

Section: Introductionmentioning

confidence: 99%

Quantum dots-based reverse phase protein microarray

Shingyoji

Gerion

Pinkel

et al. 2005

Talanta

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CdSe nanocrystals, also called quantum dots (Qdots) are a novel class of fluorophores, which have a diameter of a few nanometers and possess high quantum yield, tunable emission wavelength and photostability. They are an attractive alternative to conventional fluorescent dyes. Quantum dots can be silanized to be soluble in aqueous solution under biological conditions, and thus be used in bio-detection. In this study, we established a novel Qdot-based technology platform that can perform accurate and reproducible quantification of protein concentration in a crude cell lysate background. Protein lysates have been spiked with a target protein, and a dilution series of the cell lysate with a dynamic range of three orders of magnitude has been used for this proof-of-concept study. The dilution series has been spotted in microarray format, and protein detection has been achieved with a sensitivity that is at least comparable to standard commercial assays, which are based on horseradish peroxidase (HRP) catalyzed diaminobenzidine (DAB) chromogenesis. The data obtained through the Qdot method has shown a close linear correlation between relative fluorescence unit and relative protein concentration. The Qdot results are in almost complete agreement with data we obtained with the well-established HRP-DAB colorimetric array (R 2 =0.986). This suggests that Qdots can be used for protein quantification in microarray format, using the platform presented here.

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Protein microarrays and proteomics

Cited by 790 publications

References 47 publications

Proteome analysis of prostate cancer

Proteome analysis of prostate cancer

Application of activity-based probes to the study of enzymes involved in cancer progression

Quantum dots-based reverse phase protein microarray

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