Ultrasensitive detection of microRNAs using nanoengineered micro gold shells and laser desorption/ionization time-of-flight MS

Seo, Hyunjung; Kim, Young Ho; Kim, Jae Il; Kang, Hyunook; Jung, Woong; Yeo, Woon‐Seok

doi:10.1016/j.ab.2012.11.009

Cited by 19 publications

(1 citation statement)

References 19 publications

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“…The majority of mass spectrometry-based RNA studies used enzymatic or chemical digestion of longer RNA species prior to analysis by mass spectrometry [13]. Although analysis of miRNAs by mass spectrometry has been reported [14], previous studies have used alternative approaches to the experimental setup described here, for example direct infusion methods on a Q-TOF mass spectrometer [15] or affinity capture of a specific miRNA of interest [16]. …”

Section: Introductionmentioning

confidence: 99%

Intact MicroRNA Analysis Using High Resolution Mass Spectrometry

Kullolli

Knouf

Arampatzidou

et al. 2013

J. Am. Soc. Mass Spectrom.

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MicroRNAs (miRNAs) are small single-stranded non-coding RNAs that post-transcriptionally regulate gene expression, and play key roles in the regulation of a variety of cellular processes and in disease. New tools to analyze miRNAs will add understanding of the physiological origins and biological functions of this class of molecules. In this study we investigate the utility of high resolution mass spectrometry for the analysis of miRNAs through proof-of-concept experiments. We demonstrate the ability of mass spectrometry to resolve and separate miRNAs and corresponding 3′ variants in mixtures. The mass accuracy of the monoisotopic deprotonated peaks from various miRNAs is in the low ppm range. We compare fragmentation of miRNA by collision-induced dissociation (CID) and by higher-energy collisional dissociation (HCD) which yields similar sequence coverage from both methods but additional fragmentation by HCD versus CID. We measure the linear dynamic range, limit of detection, and limit of quantitation of miRNA loaded onto a C18 column. Lastly we explore the use of data dependent acquisition of MS/MS spectra of miRNA during online LC-MS and demonstrate that multiple charge states can be fragmented, yielding nearly full sequence coverage of miRNA on a chromatographic time scale. We conclude that high resolution mass spectrometry allows the separation and measurement of miRNAs in mixtures and a standard LC-MS setup can be adapted for online analysis of these molecules.

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Section: Introductionmentioning

confidence: 99%

Intact MicroRNA Analysis Using High Resolution Mass Spectrometry

Kullolli

Knouf

Arampatzidou

et al. 2013

J. Am. Soc. Mass Spectrom.

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Combination of Mass Signal Amplification and Isotope‐Labeled Alkanethiols for the Multiplexed Detection of miRNAs

Kang

Hong

Sung

et al. 2017

Chemistry An Asian Journal

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We report a fast and sensitive method for the multiplexed detection of miRNAs by combining mass signal amplification and isotope-labeled signal reporter molecules. In our strategy, target miRNAs are captured specifically by immobilized DNAs on gold nanoparticles (AuNPs), which carry a large number of small molecules, called amplification tags (Am-tags), as the reporter for the detection of target miRNAs. For multiplexed detection, we designed and synthesized four Am-tags containing 0, 4, 8, 12 isotopes so that they had same molecular properties but different molecular weights. By observing the mass signals of the Am-tags on AuNPs decorated along with different probe DNAs, four types of miRNAs in a sample could be easily discriminated, and the relative amounts of these miRNAs could be quantified. The practicability of our strategy was further verified by measuring the expression levels of two miRNAs in HUVECs in response to different CuSO concentrations.

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Approaches for the analysis of low molecular weight compounds with laser desorption/ionization techniques and mass spectrometry

2013

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This review summarizes various approaches for the analysis of low molecular weight (LMW) compounds by different laser desorption/ionization mass spectrometry techniques (LDI-MS). It is common to use an agent to assist the ionization, and small molecules are normally difficult to analyze by, e.g., matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) using the common matrices available today, because the latter are generally small organic compounds themselves. This often results in severe suppression of analyte peaks, or interference of the matrix and analyte signals in the low mass region. However, intrinsic properties of several LDI techniques such as high sensitivity, low sample consumption, high tolerance towards salts and solid particles, and rapid analysis have stimulated scientists to develop methods to circumvent matrix-related issues in the analysis of LMW molecules. Recent developments within this field as well as historical considerations and future prospects are presented in this review.

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Ultrasensitive detection of microRNAs using nanoengineered micro gold shells and laser desorption/ionization time-of-flight MS

Cited by 19 publications

References 19 publications

Intact MicroRNA Analysis Using High Resolution Mass Spectrometry

Intact MicroRNA Analysis Using High Resolution Mass Spectrometry

Combination of Mass Signal Amplification and Isotope‐Labeled Alkanethiols for the Multiplexed Detection of miRNAs

Approaches for the analysis of low molecular weight compounds with laser desorption/ionization techniques and mass spectrometry

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