Science Selections June 2003

Hood, Elizabeth

doi:10.1289/ehp.111-1241544

Cited by 3 publications

(4 citation statements)

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Section: Introductionmentioning

confidence: 99%

“…Developing sensitive, reliable, accurate, low cost methods for detection of cancer protein biomarkers in complex biological matrixes is a major challenge, but with high payoff of future devices for early cancer screening and point-of-care diagnosis. [1][2][3] Such methods can also lead to a better understanding of the biochemistry of disease processes and in monitoring patient's responses to therapy. 4 Various methods have been developed for protein measurement, including enzyme-linked immunosorbent assay (ELISA), [5][6][7] surface plasmon resonance (SPR), 8,9 magnetic bead-based electrochemiluminescence (ECL), 10 chemiluminescence, [11][12][13][14] fluorescence immunoassays, [15][16][17] mass spectrometry [18][19][20] and immuno-PCR.…”

Section: Introductionmentioning

confidence: 99%

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Sensitive electrochemical immunosensor for matrix metalloproteinase-3 based on single-wall carbon nanotubes

Munge

Fisher

Millord

et al. 2010

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A novel electrochemical immunosensor for the detection of matrix metalloproteinase-3 (MMP-3), a cancer biomarker protein, based on vertically aligned single-wall carbon nanotube (SWCNT) arrays is presented. Detection was based on a sandwich immunoassay consisting of horseradish peroxidase (14-16 labels) conjugated to a secondary antibody and/or a polymer bead loaded with multi-enzyme labels. Performance was optimized by effective minimization of non-specific binding (NSB) events using Bovine serum albumin (BSA), Tween-20 and optimization of the primary antibody and secondary antibody concentrations. Results provided a detection limit of 0.4 ng mL −1 (7.7 pM) for the 14-16 label sensor protocol and 4 pg mL −1 (77 fM) using a multiply enzyme labeled polymeric bead amplification strategy in 10 μL of calf serum. This immunosensor based on SWCNT arrays offers great promise for a rapid, simple, cost-effective method for clinical screening of cancer biomarkers for point-of-care diagnosis.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Sensitive electrochemical immunosensor for matrix metalloproteinase-3 based on single-wall carbon nanotubes

Munge

Fisher

Millord

et al. 2010

Analyst

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“…Genotoxic metabolites and their nucleobase adducts can be detected by separation methods such as LC−MS. , These methods are very sensitive and provide specific and detailed molecular information, but may be limited for screening by throughput, analysis time, and cost. On the other hand, alternative, relatively rapid array technologies have been very successful in genomics and proteomics and are in principle capable of many thousands of measurements on a single chip …”

mentioning

confidence: 99%

Electrochemiluminescent Arrays for Cytochrome P450-Activated Genotoxicity Screening. DNA Damage from Benzo[a]pyrene Metabolites

et al. 2007

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Arrays suitable for genotoxicity screening are reported that generate metabolites from cytochrome P450 enzymes (CYPs) in thin-film spots. Array spots containing DNA, various human cyt P450s, and electrochemiluminescence (ECL) generating metallopolymer [Ru(bpy) 2 PVP 10 ] 2+ were exposed to H 2 O 2 to activate the enzymes. ECL from all spots was visualized simultaneously using a CCD camera. Using benzo [a]pyrene as a test substrate, enzyme activity for producing DNA damage in the arrays was found in the order CYP1B1 > CYP1A2 > CYP1A1 > CYP2E1 > myoglobin, the same as the order of their metabolic activity. Thus, these arrays estimate the relative propensity of different enzymes to produce genotoxic metabolites. This is the first demonstration of ECL arrays for highthroughput in vitro genotoxicity screening.Bioactivation of xenobiotic molecules by cytochrome P450 (cyt P450, or CYP) enzymes in the human liver is a major source of genotoxicity. Metabolites formed in this way can cause damage to genetic material. 1,2 Since levels of cyt P450 enzyme expression vary dramatically in different individuals, subpopulations may be subject to varying degrees of chemical or drug toxicity. 3 Thus, knowledge of which isoforms of cyt P450 produce toxic metabolites is critical in the development of new drugs, agricultural chemicals, and other substances that impact the public.Genotoxic metabolites and their nucleobase adducts can be detected by separation methods such as LC-MS. 4,5 These methods are very sensitive and provide specific and detailed molecular information, but may be limited for screening by throughput, analysis time, and cost. On the other hand, alternative, relatively rapid array technologies have been very successful in genomics and proteomics and are in principle capable of many thousands of measurements on a single chip. 6We recently demonstrated rapid, inexpensive, voltammetric genotoxicity screening sensors assembled from films of DNA and cyt P450s in single-electrode 7 and eight-electrode formats. 8 In a two-step process, test molecules are first bioactivated by the enzymes. Then, possible adducts with DNA nucleobases are detected by voltammetry using ruthenium tris(2,2′-bipyridyl)Ru II (Ru(bpy) 3 2+ ) to catalytically oxidize the guanine bases 9 in DNA. Nucleobase adducts formed as a consequence of the enzyme reaction in the enzyme/DNA films are not detected directly, but increases in voltammetric peaks result because the adducts cause DNA * To whom correspondence should be addressed. E-mail: james.rusling@uconn.edu. † University of Connecticut. ‡ University of Connecticut Health Center. [Ru(bpy) 2 (PVP) 10 ] 2+ contains six N-bonds to Ru, and is thought to produce ECL upon reaction with guanines in DNA according to the following pathway: NIH Public AccessECL generated from adsorbed polymer films is more efficient and intense compared to ECL produced by solution species. 15 The Ru II center is oxidized by the electrode (eq 1) and subsequently oxidizes a guanine in DNA to form a guanine radical (eq 2)...

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“…Sensitive quantitative detection of protein biomarkers is critical to many areas of biomedical research and diagnostics (Smith et al, 2007), proteomics (Hood, 2003), and systems biology (Kitano, 2002). Particularly, in the systems of various diseases, the development of sensitive, rapid and low cost immunoassay method has become a central task due to the increasing need in true, in-time and expedient diagnoses (Huang et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Ultrasensitive electrochemical immunoassay based on cadmium ion-functionalized PSA@PAA nanospheres

Yin

Cui

Liu

et al. 2010

Biosensors and Bioelectronics

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Science Selections June 2003

Cited by 3 publications

References 0 publications

Sensitive electrochemical immunosensor for matrix metalloproteinase-3 based on single-wall carbon nanotubes

Sensitive electrochemical immunosensor for matrix metalloproteinase-3 based on single-wall carbon nanotubes

Electrochemiluminescent Arrays for Cytochrome P450-Activated Genotoxicity Screening. DNA Damage from Benzo[a]pyrene Metabolites

Ultrasensitive electrochemical immunoassay based on cadmium ion-functionalized PSA@PAA nanospheres

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