On-chip microfluidic dual detection of amino acid metabolism disorders using cell-free protein synthesis

Han, Jieun; Lim, Hye Jin; Park, Ju-Hwan; Han, Dong Hyun; Kim, Dong‐Myung; Park, Je‐Kyun

doi:10.1016/j.bios.2022.114936

Cited by 8 publications

(4 citation statements)

References 29 publications

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“…This open system allows not only the optimization of genes for expression , but also the testing of combinations of several genes, such as metabolic pathways − and genetic circuits, , in the laboratory. Owing to these advantages, CFPS has been used to develop methods for detecting various analytes, ranging from chemicals to biomacromolecules. − The presence of targets is transduced into the synthesis of reporter proteins, and the signal-processing steps involve amplification through transcription or translation, resulting in high sensitivities for CFPS-based assay methods. Reporters usually produce readable signals, such as fluorescence, luminescence, and electrochemistry, which can be measured using spectroscopic or amperometric detectors.…”

Section: Introductionmentioning

confidence: 99%

Sensitive Methods to Detect Single-Stranded Nucleic Acids of Food Pathogens Based on Cell-Free Protein Synthesis and Retroreflection Signal Detection

Choi,

Park,

Lee

et al. 2024

J. Agric. Food Chem.

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Cell-free protein synthesis (CFPS) has recently gained considerable attention as a new platform for developing methods to detect various molecules, ranging from small chemicals to biological macromolecules. Retroreflection has been used as an alternative signal to develop analytical methods because it can be detected by using a simple instrument comprising a white light source and a camera. Here, we report a novel reporter protein that couples the capability of CFPS and the simplicity of retroreflection signal detection. The design of the reporter was based on two pairs of protein−peptide interactions, SpyCatcher003-SpyTag003 and MDM2-PMI(N8A). MDM2-MDM2-SpyCatcher003 was decided as the reporter protein, and the two peptides, SpyTag003 and PMI(N8A), were immobilized on the surfaces of retroreflective Janus particles and microfluidic chips, respectively. The developed retroreflection signal detection system was combined with a previously reported CFPS reaction that can transduce the presence of a single-stranded nucleic acid into protein synthesis. The resulting methods were applied to detect 16S rRNAs of several foodborne pathogens. Concentration-dependent relationships were observed over a range of 10°fM to 10 2 pM, with the limits of detection being single-digit femtomolar concentrations. Considering the designability of the CFPS system for other targets, the retroreflection signal detection method will enable the development of novel methods to detect various molecules.

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Section: Introductionmentioning

confidence: 99%

Sensitive Methods to Detect Single-Stranded Nucleic Acids of Food Pathogens Based on Cell-Free Protein Synthesis and Retroreflection Signal Detection

Choi,

Park,

Lee

et al. 2024

J. Agric. Food Chem.

Self Cite

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“…It is worth noting that these techniques may address some challenges such as complex design strategies, variability among users, the requirement of an external operator, and complicated micro-valving setups often require extensive microfabrication and assembly processes. [50][51][52] Additive manufacturing (3D printing) techniques can substantially address the challenges associated with microfabrication and with microfluidic handling on a large scale. 53,54 Indeed AM can bring about outcomes comparable to traditional lithography in terms of temporal and spatial resolution, material, and mechanical properties while being cost-effective, rapid, and scalable.…”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Additive manufacturing leveraged microfluidic setup for sample to answer colorimetric detection of pathogens

Yedire,

Hosseini,

Shieh

et al. 2023

Lab Chip

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“…While typical microfluidic separators rely on additional peripherals such as pumps, syringes, microbore tubings, and connectors, capillary and finger-actuated pumps may pose challenges in recovering cells from wicking structures and separating cells due to mixing by pulsatile flow, respectively. [11][12][13][14] Alternatively, degassing-driven pumping has been adopted for equipment-free cell separation, [15][16][17] but the uncertainty about which outlet to fill first, target or waste outlets, can significantly affect separation purity and recovery. To address this issue, we employ a phase-guide structure that enables complete spatial control of wetting and filling separation channels during degassing-driven pumping, ensuring high separation performance.…”

Section: Introductionmentioning

confidence: 99%

Integrative Magneto‐Microfluidic Separation of Immune Cells Facilitates Clinical Functional Assays

Shin

Park

Lee

et al. 2023

Small

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Accurately analyzing the functional activities of natural killer (NK) cells in clinical diagnosis remains challenging due to their coupling with other immune effectors. To address this, an integrated immune cell separator is required, which necessitates a streamlined sample preparation workflow including immunological cell isolation, removal of excess red blood cells (RBCs), and buffer exchange for downstream analysis. Here, a self‐powered integrated magneto‐microfluidic cell separation (SMS) chip is presented, which outputs high‐purity target immune cells by simply inputting whole blood. The SMS chip intensifies the magnetic field gradient using an iron sphere‐filled inlet reservoir for high‐performance immuno‐magnetic cell selection and separates target cells size‐selectively using a microfluidic lattice for RBC removal and buffer exchange. In addition, the chip incorporates self‐powered microfluidic pumping through a degassed polydimethylsiloxane chip, enabling the rapid isolation of NK cells at the place of blood collection within 40 min. This chip is used to isolate NK cells from whole blood samples of hepatocellular cancer patients and healthy volunteers and examined their functional activities to identify potential abnormalities in NK cell function. The SMS chip is simple to use, rapid to sort, and requires small blood volumes, thus facilitating the use of immune cell subtypes for cell‐based diagnosis.

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On-chip microfluidic dual detection of amino acid metabolism disorders using cell-free protein synthesis

Cited by 8 publications

References 29 publications

Sensitive Methods to Detect Single-Stranded Nucleic Acids of Food Pathogens Based on Cell-Free Protein Synthesis and Retroreflection Signal Detection

Sensitive Methods to Detect Single-Stranded Nucleic Acids of Food Pathogens Based on Cell-Free Protein Synthesis and Retroreflection Signal Detection

Additive manufacturing leveraged microfluidic setup for sample to answer colorimetric detection of pathogens

Integrative Magneto‐Microfluidic Separation of Immune Cells Facilitates Clinical Functional Assays

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