Accumulation of amplified target DNAs using thiol/biotin labeling, S1 nuclease, and ferrocene–streptavidin–magnetic system and a direct detection of specific DNA signals with screen printed gold electrode

Chaumpluk, Piyasak; Kerman, Kağan; Takamura, Yuzuru; Tamiya, Eiichi

doi:10.1016/j.stam.2007.04.001

Cited by 9 publications

(9 citation statements)

References 33 publications

(41 reference statements)

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“…While these techniques provide valuable information, most of them are lack of multiplexing capability. Among those techniques that can be multiplexed, the majority of them are indirect or require further steps of gel visualization or labeling with fluorophores 21 , all of which are time and labor consuming, and are susceptible to various factors 22 .…”

Section: Introductionmentioning

confidence: 99%

A Combination of DNA-peptide Probes and Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS): A Quasi-Targeted Proteomics Approach for Multiplexed MicroRNA Quantification

Zhou

Cao

et al. 2017

Theranostics

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The distorted and unique expression of microRNAs (miRNAs) in cancer makes them an attractive source of biomarker. There is much evidence indicating that a panel of miRNAs, termed “miRNA fingerprints”, is more specific and informative than an individual miRNA as biomarker. Thus, multiplex assays for simultaneous quantification of multiple miRNAs could be more potent in clinical practice. However, current available assays normally require pre-enrichment, amplification and labeling steps, and most of them are semi-quantitative or lack of multiplexing capability. In this study, we developed a quasi-targeted proteomics assay for multiplexed miRNA quantification by a combination of DNA-peptide probes and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Specifically, the signal of target miRNAs (i.e., miR-21, miR-let7a, miR-200c, miR-125a and miR-15b) was converted into the mass response of reporter peptides by hybridization of miRNAs with DNA-peptide probes and subsequent tryptic digestion to release the peptides. After a careful optimization of conditions related to binding, conjugation, hybridization and multiple reaction monitoring (MRM) detection, the assay was validated for each miRNA and the limit of quantification (LOQ) for all the miRNAs can achieve 1 pM. Moreover, crosstalk between DNA-peptide probes in multiplex assay was sophisticatedly evaluated. Using this quasi-targeted proteomics assay, the level of target miRNAs was determined in 3 human breast cell lines and 36 matched pairs of breast tissue samples. Finally, simplex assay and qRT-PCR were also performed for a comparison. This approach grafts the strategy of targeted proteomics into miRNA quantification and may offer a new way for multiplexed miRNA profiling.

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Section: Introductionmentioning

confidence: 99%

A Combination of DNA-peptide Probes and Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS): A Quasi-Targeted Proteomics Approach for Multiplexed MicroRNA Quantification

Zhou

Cao

et al. 2017

Theranostics

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“…116 Finally, the use of Fc-(strept)avidin conjugates to study DNA hybridization events will be discussed. 115,[117][118][119] Essentially, in these systems, the interaction of biotin-DNA with Fc-strepavidin gives rise to the formation of stable complexes driven by the high formation constant of the biotin-strepavidin complex and the Fc group serves as an observable redox probe, as shown in Fig. 28.…”

Section: Fc-derivatized Proteinsmentioning

confidence: 99%

Ferrocene-peptido conjugates: From synthesis to sensory applications

et al. 2011

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The field of chemical and biological sensing is increasingly dependent on the availability of new functional materials that enhance the ability of the system to respond to chemical interactions. Organometallic bioconjugates derived from amino acids, peptides, proteins, peptide nucleic acids, and dendrimers have had a profound effect in this area and have endowed modern sensory systems with a superior performance. Owing to their fairly high stability, solubility in various solvents, and excellent redox properties, ferrocene and ferrocenyl conjugates have emerged as one of the most important classes of materials that enable direct observation of molecular interactions and as electron mediators. The low potential, reversible redox behavior of the ferrocene/ferrocenium couple is a unique property that finds widespread application in the design of sensory platforms. Currently, there is significant drive to exploit new organometallic systems, in which the presence of ferrocene acting as a redox center is critical and allows the design of highly sensitive electrochemical sensors for the sensing and recognition of a vast array of analytes.

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“…Chaumpluk et al [52] described the combination of covalently bound ferrocene-streptavidin magnetic beads (attached to biotinylated PCR products obtained by PCR with one biotinylated primer and one thiolated primer) to enrich the target sample and intercalator Hoechst 33258 for the detection of the PCR products of SSIIb and CryIA sequences from genetically modified maize. DNA from maize samples as target DNA for the PCR was obtained by means of a DNeasy Plant Mini Kit (Qiagen GmbH).…”

Section: Real Samplesmentioning

confidence: 99%

Sequence-specific electrochemical detection of nucleic acids in real samples

Duwensee¹,

Mix

Flechsig

2010

Bioanal Rev

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This paper reviews past and current developments in the field of electrochemical biosensors with a focus on the sequence-specific detection of nucleic acids in real samples. After electrochemical hybridization sensors had been first described in 1993, it took nearly a decade until some of the many proposed protocols were indeed applied to real samples like blood or tissue. Electrochemical transduction schemes used either rely on electroactive moieties such as intercalators, groove binders, covalently attached labels, and products of enzyme markers or they are completely indicator free like impedance-based detection principles. Most detection schemes require a polymerase chain reaction amplification step to allow for sufficient selectivity and sensitivity. Today, several companies develop electrochemical microarrays able to detect dozens to many thousands of sequences in a single experiment.

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Accumulation of amplified target DNAs using thiol/biotin labeling, S1 nuclease, and ferrocene–streptavidin–magnetic system and a direct detection of specific DNA signals with screen printed gold electrode

Abstract: 2007) Accumulation of amplified target DNAs using thiol/biotin labeling, S1 nuclease, and ferrocene-streptavidin-magnetic system and a direct detection of specific DNA signals with screen printed gold electrode, Science and Technology of Advanced Materials, 8:4, 323-330,

Cited by 9 publications

References 33 publications

A Combination of DNA-peptide Probes and Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS): A Quasi-Targeted Proteomics Approach for Multiplexed MicroRNA Quantification

A Combination of DNA-peptide Probes and Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS): A Quasi-Targeted Proteomics Approach for Multiplexed MicroRNA Quantification

Ferrocene-peptido conjugates: From synthesis to sensory applications

Sequence-specific electrochemical detection of nucleic acids in real samples

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