Compact Digital Immunoassay Platform Integrating ELISA with a Lateral Flow Strip DOI Creative Commons

Takuma Degawa,

Y Hori,

Masato Orikasa

et al.

Biomedicines, Journal Year: 2024, Volume and Issue: 12(11), P. 2517 - 2517

Published: Nov. 4, 2024

On-site diagnosis of infection in their early stages requires assays with high sensitivities that are compact and easy to operate out the laboratory hospital environments. However, current assay technologies fall short these requirements require highly skilled technicians set up, operate, interpret results.

Language: Английский

Single-Molecule Oxidoreductase Activity Analysis for Activity-Based Diagnosis Based on Proteoform Alterations DOI

Mayano Minoda,

Junpei Hatakeyama,

N Nagano

et al.

Journal of the American Chemical Society, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 6, 2025

We developed a single-molecule enzyme activity assay platform for NAD(P)+-dependent oxidoreductases, leveraging new NAD(P)H-responsive fluorogenic probe optimized microdevice-based fluorometric detection. This enabled the detection of activities in blood and cerebrospinal fluid (CSF), including lactate dehydrogenase, glucose-6-phosphate hexokinases. demonstrate its potential activity-based diagnosis by detecting altered populations species CSF from liver damage brain tumor patients.

Language: Английский

Citations

0
Synthetic Protein-to-DNA Input Exchange for Protease Activity Detection Using CRISPR-Cas12a DOI Creative Commons
Luca Capelli,

Federica Pedrini,

Andrea C. Di Pede

et al.

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(47), P. 18645 - 18654

Published: Nov. 14, 2024

We present a novel activity-based detection strategy for matrix metalloproteinase 2 (MMP2), critical cancer protease biomarker, leveraging mechanism responsive to the proteolytic activity of MMP2 and its integration with CRISPR-Cas12a-assisted signal amplification. designed chemical translator comprising two functional units─a peptide nucleic acid (PNA), fused together. The presents substrate MMP2, while PNA serves as output subsequent processing. This was immobilized on micrometer magnetic beads physical support an assay. incorporated into our design single-stranded DNA partially hybridized sequence bearing region complementary RNA guide CRISPR-Cas12a. target-induced nuclease Cas12a results in degradation FRET-labeled reporters amplified fluorescence signal, enabling low picomolar range, showing limit 72 pg/mL. study provides new principles broader applicability CRISPR-Cas-based biosensing.

Language: Английский

Citations

2
Thioester‐Based Coupled Fluorogenic Assays in Microdevice for the Detection of Single‐Molecule Enzyme Activities of Esterases with Specified Substrate Recognition DOI Creative Commons

T. Ukegawa,

Toru Komatsu,

Mayano Minoda

et al.

Advanced Science, Journal Year: 2023, Volume and Issue: 11(10)

Published: Dec. 22, 2023

Abstract Single‐molecule enzyme activity assay is a platform that enables the analysis of activities at single proteoform level. The limitation targetable enzymes major drawback assay, but general reported to study single‐molecule esterases based on coupled using thioesters as substrate analogues. realized by developing highly water‐soluble thiol‐reacting probes phosphonate‐substituted boron dipyrromethene (BODIPY). system detection cholinesterase in blood samples level, and it shown dissecting alterations esterase can serve an informative for activity‐based diagnosis.

Language: Английский

Citations

4
Synthetic Protein-to-DNA Input Exchange for Protease Activity Detection Using CRISPR-Cas12a DOI Creative Commons
Alessandro Bertucci, Luca Capelli,

Federica Pedrini

et al.

Published: Nov. 17, 2023

We present a novel activity-based detection strategy for matrix metalloproteinase 2 (MMP2), critical cancer protease biomarker, leveraging mechanism responsive to the proteolytic activity of MMP2 and its integration with CRISPR-Cas12a-assisted signal amplification. designed chemical translator comprising two functional units — peptide nucleic acid (PNA) fused together. The presents substrate MMP2, while PNA serves as output subsequent processing. This was immobilized on micrometric magnetic beads physical support an assay. incorporated into our design single-stranded DNA partially hybridized sequence bearing region complementary RNA guide CRISPR-Cas12a. target-induced nuclease Cas12a results in degradation FRET-labeled reporters amplified fluorescence signal, enabling low picomolar range, showing limit 72 pg/mL. study provides new principles broader applicability CRISPR-Cas-based biosensing.

Language: Английский

Citations

1
Synthetic Protein-to-DNA Input Exchange for Protease Activity Detection Using CRISPR-Cas12a DOI Creative Commons
Luca Capelli,

Federica Pedrini,

Andrea C. Di Pede

et al.

Published: Nov. 29, 2023

We present a novel activity-based detection strategy for matrix metalloproteinase 2 (MMP2), critical cancer protease biomarker, leveraging mechanism responsive to the proteolytic activity of MMP2 and its integration with CRISPR-Cas12a-assisted signal amplification. designed chemical translator comprising two functional units — peptide nucleic acid (PNA) fused together. The presents substrate MMP2, while PNA serves as output subsequent processing. This was immobilized on micrometric magnetic beads physical support an assay. incorporated into our design single-stranded DNA partially hybridized sequence bearing region complementary RNA guide CRISPR-Cas12a. target-induced nuclease Cas12a results in degradation FRET-labeled reporters amplified fluorescence signal, enabling low picomolar range, showing limit 72 pg/mL. study provides new principles broader applicability CRISPR-Cas-based biosensing.

Language: Английский

Citations

1
Exploring fluoropolymers for fabrication of femtoliter chamber arrays used in digital bioanalysis DOI Creative Commons
Jun Ando,

Kazue Murai,

Makiko Mori

et al.

Scientific Reports, Journal Year: 2024, Volume and Issue: 14(1)

Published: May 20, 2024

The global supply of fluoropolymers and fluorinated solvents is decreasing due to environmental concerns regarding polyfluoroalkyl substances. CYTOP has been used for decades primarily as a component femtoliter chamber array digital bioanalysis; however, its recently become scarce, increasing the urgency fabricating using alternative materials. In this study, we investigated feasibility four types in stable candidate substitutes verified their applicability bioanalysis. Among these candidates, Fluorine Sealant emerged viable option arrays conventional photolithography process. To validate efficacy, performed various bioanalysis FP-A-based with model enzymes such CRISPR-Cas, horseradish peroxidase, β-galactosidase. results demonstrated similar performance that CYTOP, highlighting broader utility FP-A Our findings underscore potential enhance versatility foster ongoing advancement innovative diagnostic technologies.

Language: Английский

Citations

0
Compact Digital Immunoassay Platform Integrating ELISA with a Lateral Flow Strip DOI Creative Commons

Takuma Degawa,

Y Hori,

Masato Orikasa

et al.

Biomedicines, Journal Year: 2024, Volume and Issue: 12(11), P. 2517 - 2517

Published: Nov. 4, 2024

On-site diagnosis of infection in their early stages requires assays with high sensitivities that are compact and easy to operate out the laboratory hospital environments. However, current assay technologies fall short these requirements require highly skilled technicians set up, operate, interpret results.

Language: Английский

Citations

0