An open-space microfluidic chip with fluid walls for online detection of VEGF <i>via</i> rolling circle amplification

Feng, Shuo; Mao, Sifeng; Dou, Jinxin; Li, Weiwei; Li, Haifang; Lin, Jin‐Ming

doi:10.1039/c9sc02974e

Cited by 23 publications

(23 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The dataset about I p vs the logarithmic c target , Log( c target ), could be fitted into a linear calibration curve [ I p = (99.3 ± 12.0) Log( c target ) + (211.8 ± 13.1)] with a squared regression of 0.993 (Figure C, inset). The limit of detection (LOD) at a signal-to-noise ratio of 3 was gauged to be 0.6 pg·mL –1 , which is commeasurable to the performance of the industrially well-renowned SULFO-TAG in Meso Scale Discovery ECL imagers, and even beats some cases involving proof-of-concept nanostructures. , Noticeably, such abzyme-liked conjugate of Ab 2 -biotin/strp-Hb ZnPPIX achieved a wide dynamic scope over 5 orders of magnitude.…”

Section: Resultsmentioning

confidence: 96%

“…The limit of detection (LOD) at a signal-to-noise ratio of 3 was gauged to be 0.6 pg•mL −1 , which is commeasurable to the performance of the industrially wellrenowned SULFO-TAG in Meso Scale Discovery ECL imagers, 56 and even beats some cases involving proof-ofconcept nanostructures. 57,58 Noticeably, such abzyme-liked conjugate of Ab 2 -biotin/strp-Hb ZnPPIX achieved a wide dynamic scope over 5 orders of magnitude.…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

See 1 more Smart Citation

Electrochemiluminescence-Repurposed Abiological Catalysts in Full Protein Tag for Ultrasensitive Immunoassay

et al. 2020

View full text Add to dashboard Cite

Being announced as one of the “2019 Top Ten Emerging Technologies in Chemistry” by IUPAC, the directed evolution of artificial metalloenzymes has led to a broad scope of abiotic processes. Here, inspired by those key proteins in bioluminescence, a rudimentary expression of bio-electrochemiluminescent (ECL) macromolecules was achieved via the complexation of zinc proto-porphyrin IX (ZnPPIX) within apo-hemoglobin (apo-Hb). A high-yield monochromic irradiation at 644 nm could be provoked potentiostatically from the reconstituted holo-HbZnPPIX in solutions. Its secondary structure integrity was elucidated by UV and circular dichroism spectrometry, while voltammetry-hyphenated surface plasmon resonance authenticated its ligation conservativeness in electrical fields. Further conjugation with streptavidin rendered a homogeneous Janus fusion of both receptor and reporter domains, enabling a new abiological catalyst-linked ECL bioassay. On the other hand, singular ZnPPIX inside each tetrameric subunit of Hb accomplished an overall signal amplification without the bother of luminogenic heterojunctions. This pH-tolerant and non-photobleaching optics was essentialized to be the unique configuration interaction between Zn and O2, by which the direct electrochemistry of proteins catalyzed the transient progression of O2 → O2 ·– → O2* + hυ selectively. Such principle was implemented as a signal-on strategy for the determination of a characteristic cancer biomarker, the vascular endothelial growth factor, resulting in competent performance at a low detection limit of 0.6 pg·mL–1 and a wide calibration range along with good stability and reliability in real practices. This simple mutation repurposed the O2-transport Hb in the erythrocytes of almost all vertebrates into a cluster of oxidoreductases with intrinsic ECL activity, which would enrich the chromophore library. More importantly, its genetically engineered variants may come in handy in biomedical diagnosis and visual electrophysiology.

show abstract

Section: Resultsmentioning

confidence: 96%

Section: ■ Experimental Sectionmentioning

confidence: 99%

Electrochemiluminescence-Repurposed Abiological Catalysts in Full Protein Tag for Ultrasensitive Immunoassay

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Typical studies have been exemplified to demonstrate the approaches that microfluidics can be exploited for the analysis of these biospecies, followed by the most recent advances in translating the analysis of biospecies into practical applications. As illustrated in Figure , microfluidic devices can be made of various materials such as inorganic materials, polymers, and paper, according to different applications. , Integration of microdevices with a variety of chip designs will facilitate the promising function of them, especially when integrating with nanomaterials. For the analysis of biospecies, a series of detection methods including optical techniques, electrochemical, or electrical approaches can be utilized, in combination with microfluidic platforms. − Therefore, with the rapid and sensitive detection of various biospecies including proteins, nucleic acids, cells, pathogen, etc., microfluidic biosensors exhibit promising applications in different fields, e.g., diagnosis of disease, food safety control, and environmental monitoring.…”

Section: Introductionmentioning

confidence: 99%

Microfluidics-Based Sensing of Biospecies

Xing

Zhao

Cheng

et al. 2020

ACS Appl. Bio Mater.

View full text Add to dashboard Cite

Over the past decades, microfluidic devices based on many advanced techniques have aroused widespread attention in the fields of chemical, biological, and analytical applications. Integration of microdevices with a variety of chip designs will facilitate promising functionality. Notably, the combination of microfluidics with functional nanomaterials may provide creative ideas to achieve rapid and sensitive detection of various biospecies. In this review, focused on the microfluids and microdevices in terms of their fabrication, integration, and functions, we summarize the up-to-date developments in microfluidics-based analysis of biospecies, where biomarkers, small molecules, cells, and pathogens as representative biospecies have been explored in-depth. The promising applications of microfluidic biosensors including clinical diagnosis, food safety control, and environmental monitoring are also discussed. This review aims to highlight the importance of microfluidics-based biosensors in achieving high throughput, highly sensitive, and low-cost analysis and to promote microfluidics toward a wider range of applications.

show abstract

“…12 Lin et al also recently developed an open-space microfluidic chip with fluid walls for the online detection of VEGF (vascular endothelial growth factor) via RCA. 13 However, exotic cofactors (such as enzymes or metal ions) necessary for these studies impede their further utility inside living cells. Moreover, the inherent flaws in amplification strategies may result in reduced specificity because any accumulation of parasitic amplicons or nonspecific may yield interfering signals.…”

mentioning

confidence: 99%

“…For example, Wang et al reported on gold nanoparticle-based DNAzyme probes for intracellular miRNA detection. , Peng et al described telomerase-catalyzed FRET ratio flares with signal amplification for intracellular miRNA imaging . Lin et al also recently developed an open-space microfluidic chip with fluid walls for the online detection of VEGF (vascular endothelial growth factor) via RCA . However, exotic cofactors (such as enzymes or metal ions) necessary for these studies impede their further utility inside living cells.…”

mentioning

confidence: 99%

Construction of Dual-Color Probes with Target-Triggered Signal Amplification for In Situ Single-Molecule Imaging of MicroRNA

Liu

et al. 2020

ACS Nano

View full text Add to dashboard Cite

The in vitro detection of low abundance biomolecules via nonenzymatic signal amplification is an attractive strategy. However, it remains a challenge to monitor targets of interest in situ in living cells by low-background interference and visualized enzyme-free signal amplification strategies. Taking advantage of the single-molecule imaging and dynamic DNA nanotechnologies, we have achieved the target-triggered self-assembly of nanostructure-based dual-color fluorescent probes (NDFPs) by an enzyme-free toehold-mediated strand displacement cascade. NDFPs will facilitate the simple and visualized monitoring of microRNA (miRNA) at the femtomolar level. The recycled miRNA can be considered as the catalyst for the assembly of multiple H1/H2 duplexes. This generated the fluorescence signal of the enhanced target expression, indicating both in vitro and in vivo signal-amplified imaging. Moreover, the NDFPs improved the measurement accuracy by dual-color colocalization imaging to greatly avoid false-positive signals and enabled the successful in situ imaging of miRNA in living cells in real time. This work provides a strategy to visually monitor and study the integration of signal amplification detection and single-molecule imaging. NDFPs may be an important step toward the enzyme-free amplified monitoring and imaging of various biomolecules in living cells at the single-molecule level.

show abstract

An open-space microfluidic chip with fluid walls for online detection of VEGF via rolling circle amplification

Abstract: We report an open-space microfluidic chip with fluid walls, integrating functions of cell culture and online detection of secreted proteins controlled by the interfacial tension value.

Cited by 23 publications

References 44 publications

Electrochemiluminescence-Repurposed Abiological Catalysts in Full Protein Tag for Ultrasensitive Immunoassay

Electrochemiluminescence-Repurposed Abiological Catalysts in Full Protein Tag for Ultrasensitive Immunoassay

Microfluidics-Based Sensing of Biospecies

Construction of Dual-Color Probes with Target-Triggered Signal Amplification for In Situ Single-Molecule Imaging of MicroRNA

Contact Info

Product

Resources

About