New functional protein measurement technology could accelerate drug discovery research

a Schematic illustration of the CycMIST process for analyzing multiple proteins using the MIST microsphere array. b Distribution of the number of oligo-coated DNA microbeads on each 75 μm × 75 μm area of ​​the MIST array corresponding to the PDMS microwell, n = 3 independent MIST array. c Distribution of the number of the same type of oligo DNA-coated microspheres in the same MIST array, n = 5 independent MIST arrays. d Characterization of CycMIST sensitivity by varying the concentrations of 50 biotinylated complementary oligo DNAs on the MIST array, n = 10 independent experiments. This is the same procedure in experiments to detect single-celled proteins, except for cellular loading and conjugate binding. e Consistency of fluorescence intensities for 4 decoding cycles and for 3 fluorescent color dyes (Alexa Fluor 488, Cy3 and Cy5), n = 5 independent experiments. f Exemplary images of a multiplex analysis of 50 proteins from a single CycMIST cell and images from 4 decoding cycles. The grayscale images are the result of protein detection, and the color images are the decoding cycles from cycle 1 to cycle 4. The bottom panel is the magnified images from the squares in the top panel. Scale band: 20 μm (panel up); 5 μm (bottom panel). Data are presented as means ± SD for more than three independent experiments, and error bars are within the character size if not shown. The term (arb. Units) is abbreviated to arbitrary units. Credit: Nature Communications (2022). DOI: 10.1038 / s41467-022-31336-x “width =” 800 “height =” 530 “/>

Review of CycMIST technology for analysis of a single-celled functional proteome. a Schematic illustration of the CycMIST process for analyzing multiple proteins using the MIST microsphere array. b Distribution of the number of oligo-coated DNA microspheres in each 75 μm × 75 μm region of the MIST array corresponding to the PDMS microwell, n = 3 independent MIST array. ° C Distribution of the number of the same type of oligo-DNA-coated microspheres on the same MIST array, n = 5 independent MIST array. e Characterization of CycMIST sensitivity by changing the concentrations of 50 biotinylated complementary oligo DNAs on the MIST array, n = 10 independent experiments. This is the same procedure in experiments to detect single-celled proteins, except for cellular loading and conjugate binding. e Consistency of fluorescence intensities for 4 decoding cycles and for 3 fluorescent color dyes (Alexa Fluor 488, Cy3 and Cy5), n = 5 independent experiments. is Exemplary images of multiplex analysis of 50 proteins from a single CycMIST cell and images with 4 decoding cycles. The grayscale images are the result of protein detection, and the color images are the decoding cycles from cycle 1 to cycle 4. The bottom panel is the magnified images from the squares in the top panel. Scale band: 20 μm (panel up); 5 μm (bottom panel). Data are presented as means ± SD for more than three independent experiments, and error bars are within the character size if not shown. The term (arb. Units) is abbreviated to arbitrary units. credit: Natural communications (2022). DOI: 10.1038 / s41467-022-31336-x

A new biomedical research tool that allows scientists to measure hundreds of functional proteins in a single cell may offer new insights into cellular machines. Led by Jun Wang, an associate professor of biomedical engineering at Stony Brook University, this microchip analysis – called single-cell cyclic multiplex technology in situ (CycMIST) – could help advance advances in areas such as molecular diagnostics and drug discovery. Details of the cyclic microchip analysis method are published in Natural communications.

While newer single-cell omics technologies (ie, genomics, transcriptomics, etc.) have revolutionized the study of complex biological and cellular systems, and scientists can analyze the genomic sequences of individual cells, these technologies do not apply to proteins because are not amplified as DNA. Thus, protein analysis in single cells did not lead to large-scale experiments. Because proteins are cellular functions and biomarkers for cell types and disease diagnosis, additional single-cell analysis is needed.

“The CycMIST test allows for a comprehensive assessment of cell function and physiological status by examining 100 times more protein species than conventional immunofluorescence staining, a feature unmatched by any other similar technology,” explains Liwei Young, lead author of the study and postdoctoral fellow. in the research team of Wang and Multiplex Biotechnology Laboratory.

Wang, who is affiliated with the Renaissance School of Medicine and the Stony Brook Cancer Center, and colleagues demonstrated CycMIST by detecting 182 proteins that include surface markers, neuronal function proteins, neurodegeneration markers, signaling pathway proteins, and transcription factors. They used a model of Alzheimer’s disease (AD) in mice to validate the technology and method.

By analyzing 182 proteins with CycMIST, they were able to perform a functional protein assay that revealed the deep heterogeneity of brain cells, distinguished AD markers, and identified the mechanisms of AD pathogenesis.

With this detailed way of detecting proteins in the AD model, the team suggests that such a functional protein assay may be promising for new drug targets for AD for which there is still no effective treatment. And they provide a landscape of potential drug targets at the cellular level from the CycMIST protein assay.

The authors believe that CycMIST may also have huge potential for commercialization.

They say that before this model of testing with CycMIST, researchers were able to measure and know only one peak of protein species in a cell. But this new approach allows scientists to identify and know the actions of each aspect of the cell and therefore can potentially identify whether the cell is in a disease state or not – the first step in a possible way to diagnose the disease by analyzing a single protein cell. And compared to standard approaches such as flow cytometry, their CycMIST approach can analyze 10 times the amount of protein at the single cell level.

The researchers also suggest that the analysis of the cyclic microchip is portable, inexpensive, and can be adapted to any existing fluorescence microscope, which are additional reasons for its marketability if it proves effective with subsequent experimentation.


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More info:
Liwei Yang et al, Cyclic microchip analysis to measure hundreds of functional proteins in single neurons, Natural communications (2022). DOI: 10.1038 / s41467-022-31336-x

Provided by Stony Brook University

Quote: New Technology for Measuring Functional Proteins Can Accelerate Drug Detection Research (2022, June 29), retrieved June 29, 2022 from https://phys.org/news/2022-06-functional-protein-technology -advance-drug.html

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