Journal: Analytical chemistry

Article Title: Protein and Proteome Measurements with Microfluidic Chips

doi: 10.1021/acs.analchem.9b04711

Figure Lengend Snippet: Schematic representation (a) and experimental setup (b) of a piezo-ring-on-chip microfluidic device for depositing sample on a MALDI-MS target plate in small 40 nL volumes. Reprinted with permission from Tsao, C.-W.; Lei, I. C.; Chen, P.-Y.; Yang, Y.-L. Analyst 2018, 143, 981–988 (ref 20). Copyright 2018 Centre National de la Recherche Scientifique (CNRS) and The Royal Society of Chemistry.

Article Snippet: 47 Recently, a microfluidic single-cell resolution Western blot device and method capable of capturing a panel of 12 proteins from ER+ and HER2+ circulating breast cancer cells was reported by Sinkala et al. 49 The microfluidic workflow included CTC enrichment from 2 to 4 mL of blood using a commercial size and deformability-selective microfluidic device (Vortex Biosciences), Hoechst/nuclear staining for visualizing the putative CTCs, single-cell CTC deposition into a microwell (50 μ m diameter), and CTC lysis in the well, followed by PAGE separation (within seconds), covalent immobilization of proteins to the gel, and Western immunostaining of the panel of 12 proteins.

Techniques:

Journal: Analytical chemistry

Article Title: Protein and Proteome Measurements with Microfluidic Chips

doi: 10.1021/acs.analchem.9b04711

Figure Lengend Snippet: Integrated platform composed of a microfluidic device and a His/Myc-double-tagged ORF library spotted on glass for monitoring the parallel expressions and phosphorylation of thousands of proteins. One assay unit contains DNA and protein chambers isolated by valves. (a) The microfluidic device. (b) Following incubation with a mammalian reticulocyte lysate, the target proteins are expressed in the DNA chambers, diffused, and immobilized in the protein chambers via a His tag. (c) Proteins expressed in mammalian cell lysates; the enzymes that have autophosphorylation activity are expected to be functional and undergo autophosphorylation during expression. (d, e) The proteins and the p-Tyr proteins are both quantified in situ using Cy3-coupled anti-Myc antibodies and Cy5-coupled antiphosphorylated-Tyr antibodies, respectively. Adapted with permission from Nevenzal, H.; Noach-Hirsh, M.; Skornik-Bustan, O.; Brio, L.; Barbiro-Michaely, E.; Glick, Y.; Avrahami, D.; Lahmi, R.; Tzur, A.; Gerber, D. Commun. Biol. 2019, 2, 42 (ref 27). Copyright 2019 granted underCreative Commons CC BY license (https://creativecommons.org/licenses/by/4.0/).

Article Snippet: 47 Recently, a microfluidic single-cell resolution Western blot device and method capable of capturing a panel of 12 proteins from ER+ and HER2+ circulating breast cancer cells was reported by Sinkala et al. 49 The microfluidic workflow included CTC enrichment from 2 to 4 mL of blood using a commercial size and deformability-selective microfluidic device (Vortex Biosciences), Hoechst/nuclear staining for visualizing the putative CTCs, single-cell CTC deposition into a microwell (50 μ m diameter), and CTC lysis in the well, followed by PAGE separation (within seconds), covalent immobilization of proteins to the gel, and Western immunostaining of the panel of 12 proteins.

Techniques: Isolation, Incubation, Activity Assay, Functional Assay, Expressing, In Situ

Journal: Analytical chemistry

Article Title: Protein and Proteome Measurements with Microfluidic Chips

doi: 10.1021/acs.analchem.9b04711

Figure Lengend Snippet: Schematic representation of a microfluidic chip for performing HDX of proteins on subsecond time scales. Reprinted from Svejdal, R. R.; Dickinson, E. R.; Sticker, D.; Kutter, J. P.; Rand, K. D. Anal. Chem. 2019, 91, 1309–1317 (ref 29). Copyright 2018 American Chemical Society.

Article Snippet: 47 Recently, a microfluidic single-cell resolution Western blot device and method capable of capturing a panel of 12 proteins from ER+ and HER2+ circulating breast cancer cells was reported by Sinkala et al. 49 The microfluidic workflow included CTC enrichment from 2 to 4 mL of blood using a commercial size and deformability-selective microfluidic device (Vortex Biosciences), Hoechst/nuclear staining for visualizing the putative CTCs, single-cell CTC deposition into a microwell (50 μ m diameter), and CTC lysis in the well, followed by PAGE separation (within seconds), covalent immobilization of proteins to the gel, and Western immunostaining of the panel of 12 proteins.

Techniques:

Journal: Analytical chemistry

Article Title: Protein and Proteome Measurements with Microfluidic Chips

doi: 10.1021/acs.analchem.9b04711

Figure Lengend Snippet: Microfluidic IMER chip for proteolytic digestion with trypsin adsorbed on the porous surface of PDMS channels. (a) Microfluidic chip. (b) Multiplexed design of the IMER channels. (c) Serpentine IMER channels. Reproduced with permission from Kecskemeti, A.; Nagy, C.; Csosz, E.; Kallo, G.; Gaspar, A. Prot. Clin. Appl. 2017, 11, 11–12, 1700055 (ref 39). Copyright 2017 John Wiley and Sons.

Article Snippet: 47 Recently, a microfluidic single-cell resolution Western blot device and method capable of capturing a panel of 12 proteins from ER+ and HER2+ circulating breast cancer cells was reported by Sinkala et al. 49 The microfluidic workflow included CTC enrichment from 2 to 4 mL of blood using a commercial size and deformability-selective microfluidic device (Vortex Biosciences), Hoechst/nuclear staining for visualizing the putative CTCs, single-cell CTC deposition into a microwell (50 μ m diameter), and CTC lysis in the well, followed by PAGE separation (within seconds), covalent immobilization of proteins to the gel, and Western immunostaining of the panel of 12 proteins.

Techniques:

Journal: Analytical chemistry

Article Title: Protein and Proteome Measurements with Microfluidic Chips

doi: 10.1021/acs.analchem.9b04711

Figure Lengend Snippet: Microfluidic device for desalting protein samples with a silica isoporous membrane. (a) Microfluidic assembly. (b) Serpentine PMMA channel for sample delivery. (c) Representation of the desalting process through the membrane. Reproduced from Wu, W.; Zhang, D.; Chen, K.; Zhou, P.; Zhao, M.; Qiao, L.; Su, B. Anal. Chem. 2018, 90, 14395–14401 (ref 42). Copyright 2018 American Chemical Society.

Article Snippet: 47 Recently, a microfluidic single-cell resolution Western blot device and method capable of capturing a panel of 12 proteins from ER+ and HER2+ circulating breast cancer cells was reported by Sinkala et al. 49 The microfluidic workflow included CTC enrichment from 2 to 4 mL of blood using a commercial size and deformability-selective microfluidic device (Vortex Biosciences), Hoechst/nuclear staining for visualizing the putative CTCs, single-cell CTC deposition into a microwell (50 μ m diameter), and CTC lysis in the well, followed by PAGE separation (within seconds), covalent immobilization of proteins to the gel, and Western immunostaining of the panel of 12 proteins.

Techniques:

Journal: Analytical chemistry

Article Title: Protein and Proteome Measurements with Microfluidic Chips

doi: 10.1021/acs.analchem.9b04711

Figure Lengend Snippet: Workflow for selecting THP-1 cells with a microfluidic cell sorter device to test the ability to perform proteomic profiling of rare CTCs. Cell collection was performed in tubes with various coatings to enable different processing procedures prior to LC-MS/MS analysis: Group 1, 1000 cells in a BSA-coated tube digested with medium; Group 2, 1000 cells in a BSA-coated tube with the medium removed; Group 3, 1000 cells in a hydrophilic-coated tube with the medium removed; Group 4, 100 cells in a BSA-coated tube digested with medium; Group 5, 100 cells in a BSA-coated tube with the medium removed. Reprinted with permission from Kasuga, K.; Katoh, Y.; Nagase, K.; Igarashi, K. Proteomics 2017, 17, 1600420 (ref 52). Copyright 2017 John Wiley and Sons.

Article Snippet: 47 Recently, a microfluidic single-cell resolution Western blot device and method capable of capturing a panel of 12 proteins from ER+ and HER2+ circulating breast cancer cells was reported by Sinkala et al. 49 The microfluidic workflow included CTC enrichment from 2 to 4 mL of blood using a commercial size and deformability-selective microfluidic device (Vortex Biosciences), Hoechst/nuclear staining for visualizing the putative CTCs, single-cell CTC deposition into a microwell (50 μ m diameter), and CTC lysis in the well, followed by PAGE separation (within seconds), covalent immobilization of proteins to the gel, and Western immunostaining of the panel of 12 proteins.

Techniques: Liquid Chromatography with Mass Spectroscopy

Journal: Analytical chemistry

Article Title: Protein and Proteome Measurements with Microfluidic Chips

doi: 10.1021/acs.analchem.9b04711

Figure Lengend Snippet: Schematic diagram of a multiplexed immunoassay microfluidic chip for analyzing cell-secreted cytokines. The chip contains 16 units, each comprising a cell culture chamber, transfer pump for cell culture media, and a chamber with 10 antibody capture spots for analyzing the secreted cytokines in the cell supernatant. The control layer (red) modulates the fluid flow (blue) in the cell/immunoassay layer. Reprinted with permission from Kaestli, A. J.; Junkin, M.; Tay, S. Lab Chip 2017, 17, 4124–4133 (ref 61). Copyright 2017 The Royal Society of Chemistry.

Article Snippet: 47 Recently, a microfluidic single-cell resolution Western blot device and method capable of capturing a panel of 12 proteins from ER+ and HER2+ circulating breast cancer cells was reported by Sinkala et al. 49 The microfluidic workflow included CTC enrichment from 2 to 4 mL of blood using a commercial size and deformability-selective microfluidic device (Vortex Biosciences), Hoechst/nuclear staining for visualizing the putative CTCs, single-cell CTC deposition into a microwell (50 μ m diameter), and CTC lysis in the well, followed by PAGE separation (within seconds), covalent immobilization of proteins to the gel, and Western immunostaining of the panel of 12 proteins.

Techniques: Cell Culture

Journal: Analytical chemistry

Article Title: Protein and Proteome Measurements with Microfluidic Chips

doi: 10.1021/acs.analchem.9b04711

Figure Lengend Snippet: Microfluidic chip incorporating a reactor that enables cell culture, stimulation, and cell recovery for monitoring proteome-level fast cellular responses by mass spectrometry (a). COMSOL simulation of transversal delivery of a stimulant solution to the cell chamber (b–d). Reprinted with permission from Lazar, I. M.; Deng, J.; Stremler, M. A.; Ahuja, S. Microsystems & Nanoengineering 2019, 5, 7 (ref 68). Copyright granted under Creative Commons CC BY license (https://creativecommons.org/licenses/by/4.0/).

Article Snippet: 47 Recently, a microfluidic single-cell resolution Western blot device and method capable of capturing a panel of 12 proteins from ER+ and HER2+ circulating breast cancer cells was reported by Sinkala et al. 49 The microfluidic workflow included CTC enrichment from 2 to 4 mL of blood using a commercial size and deformability-selective microfluidic device (Vortex Biosciences), Hoechst/nuclear staining for visualizing the putative CTCs, single-cell CTC deposition into a microwell (50 μ m diameter), and CTC lysis in the well, followed by PAGE separation (within seconds), covalent immobilization of proteins to the gel, and Western immunostaining of the panel of 12 proteins.

Techniques: Cell Culture, Mass Spectrometry