A highly efficient and versatile microchip capillary electrophoresis method for DNA separation using gold nanoparticle as a tag

Cao, Wei; Chen, Lei; Fu, Youjian; Tan, Zhenyi; Qu, Bin

doi:10.1002/jssc.201000755

Cited by 19 publications

(13 citation statements)

References 46 publications

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“…However, it seems that C 4 D has not yet been tried for such CE application. Nor did other electrochemical methods find use in CE of individual AuNPs (but AuNPs utilized as a tag to enhance resolution between other types of analytes ). In its turn, ICP‐MS appears to be of great suitability for detecting inorganic NPs by the virtue of a highly sensitive and specific signal afforded for a great many core/shell metals, importantly, not only in a standalone configuration but also as the on‐line coupling with separation techniques such as field‐flow fractionation or LC .…”

Section: Introductionmentioning

confidence: 99%

Comparison of detection techniques for capillary electrophoresis analysis of gold nanoparticles

et al. 2015

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As metallic nanoparticles are growing in importance as analytes in CE, increases an interest in appropriate detection methods for their quantification in various samples. For gold nanoparticles (AuNPs), the most common UV detection poses intricacy of inadequate sensitivity that hinders the applicability of CE. With the objective of resolving this challenge, UV detection was compared with C(4) D and ICP-MS as alternative modes of detection for AuNPs. A C(4) D detector, applied under pressure-driven conditions, exhibited better sensitivity than a UV detector. However, C(4) D turned to be unsatisfactory to differentiate the signal of AuNPs at common CE conditions despite varying the nature of BGE and detection conditions. Due to intrinsic sensitivity and low background levels typical to Au, ICP-MS greatly surpasses UV detection. After optimization trials, CE-ICP-MS gained the LOD of AuNPs as low as 2 × 10(-15) M, as well as an excellent performance in terms of signal stability and linearity. Also importantly, the optimized BGE appears to be well matched to explore the behavior of AuNPs in biologically relevant systems. This was demonstrated by probing the interaction between AuNPs and the main blood-transporting protein, HSA.

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Section: Introductionmentioning

confidence: 99%

Comparison of detection techniques for capillary electrophoresis analysis of gold nanoparticles

et al. 2015

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“…For instance, a neutral or a positively charged drag‐tag would slow down the DNAs of different lengths by different amounts (the shorter the DNA, the larger the influence of the drag‐tag), thus leading to separation. Experimental tests of this idea have been conducted using different types of drag‐tags, for example, a globular protein streptavidin ; unfolded peptides, both neutral and charged ; branched polypeptoids ; micelles serving as transiently bound drag‐tags ; and gold nanoparticles . To date, ELFSE has been used to separate ssDNA of up to 265 bases with single‐base resolution , but further progress requires both experimental effort and better theoretical understanding.…”

Section: Introductionmentioning

confidence: 99%

Theory of end‐labeled free‐solution electrophoresis: Is the end effect important?

Chubynsky

Slater

2014

Electrophoresis

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In the theory of free-solution electrophoresis of a polyelectrolyte (such as the DNA) conjugated with a "drag-tag," the conjugate is divided into segments of equal hydrodynamic friction and its electrophoretic mobility is calculated as a weighted average of the mobilities of individual segments. If all the weights are assumed equal, then for an electrically neutral drag-tag, the elution time t is predicted to depend linearly on the inverse DNA length 1/M. While it is well-known that the equal-weights assumption is approximate and in reality the weights increase toward the ends, this "end effect" has been assumed to be small, since in experiments the t(1/M) dependence seems to be nearly perfectly linear. We challenge this assumption pointing out that some experimental linear fits do not extrapolate to the free (i.e. untagged) DNA elution time in the limit 1/M→0, indicating nonlinearity outside the fitting range. We show that a theory for a flexible polymer taking the end effect into account produces a nonlinear curve that, however, can be fitted with a straight line over a limited range of 1/M typical of experiments, but with a "wrong" intercept, which explains the experimental results without additional assumptions. We also study the influence of the flexibilities of the charged and neutral parts.

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“…Nanomaterials offer properties such as large surface area-to-volume ratio and relative easy functionalization for preconcentration and separation steps (e.g., by coupling to antibodies or other biomolecules). Also, nanomaterials have been used as buffer additives and as stationary phases in microchip electrochromatography [ 26 ]. Furthermore, nanomaterials offer catalytic properties and capability to act as electron-transfer mediators to improve analytical reaction in microfluidic methods [ 27 ].…”

Section: Discussionmentioning

confidence: 99%

A microfluidic multiplex proteomic immunoassay device for translational research

et al. 2015

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ObjectiveMicrofluidic technology has the potential to miniaturize and automate complex laboratory procedures. The objective of this study was to assess a microfluidic immunoassay device, Simple Plex, which simultaneously measured IL-1β, TNF-α, IL-6, and IL-10 in serum samples. This assessment is important to understanding the potentials of this microfluidic device as a valuable tool in translational research efforts.MethodsWe studied the operational characteristics of Simple Plex, and compared to other immunoassay systems including bead-based (i.e., Bio-Plex® from Bio-Rad) and planar micro-spot based (i.e., Multi-Array from Meso Scale Discovery) multiplex assays. We determined imprecisions for each of the Simple Plex assays and evaluated the ability of Simple Plex to detect IL-1β, TNF-α, IL-6, and IL-10 in serum samples.ResultsSimple Plex assays required 25 µL serum, and 1.5 h to run 16 samples per cartridge per instrument. Assay imprecisions, evaluated by measurement of 6 replicates in duplicate from a serum pool using three different cartridges, were less than 10 % for all 4 cytokine protein biomarkers, comparable to the imprecisions of traditional ELISAs. The Simple Plex assays were able to detect 32, 95, 97, and 100 % [i.e., percentages of the results within the respective analytical measurement ranges (AMRs)] of IL-1β, TNF-α, IL-6, and IL-10, respectively, in 66 serum samples.ConclusionsSimple Plex is a microfluidic multiplex immunoassay device that offers miniaturized, and automated analysis of protein biomarkers. Microfluidic devices such as Simple Plex represent a promising platform to be used in translational research to measure protein biomarkers in real clinical samples.

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A highly efficient and versatile microchip capillary electrophoresis method for DNA separation using gold nanoparticle as a tag

Cited by 19 publications

References 46 publications

Comparison of detection techniques for capillary electrophoresis analysis of gold nanoparticles

Comparison of detection techniques for capillary electrophoresis analysis of gold nanoparticles

Theory of end‐labeled free‐solution electrophoresis: Is the end effect important?

A microfluidic multiplex proteomic immunoassay device for translational research

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