Journal article
Squeezable Hydrogel Microparticles for Single Extracellular Vesicle Protein Profiling
Small (Weinheim an der Bergstrasse, Germany), e2407809
29 Oct 2024
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Abstract Extracellular vesicles (EVs) are promising for molecular diagnostics, but current analyses are limited by the rarity and compositional heterogeneity of EV protein expression. Therefore, single EV profiling methods require high sensitivity, multiplexing, and throughput to address these issues. Here a single EV analysis technique that utilizes squeezable methacrylated hyaluronic acid hydrogel microparticles (MHPs) is described as a scaffold to immobilize EVs and perform an integrated rolling circle amplification (RCA) assay for an ultra‐sensitive and multiplex analysis of single EV proteins. EVs are prepared into MHPs in a high‐throughput manner with droplet microfluidics and optimally labeled with antibody‐oligonucleotide conjugates in MHPs without steric limitations. By designing MHPs with high compressibility, single EV protein signals are amplified as RCA products that can be aligned on the same plane by physically squeezing MHPs and visualized with low magnification. This method provides a simple and scalable single EV imaging analysis pipeline for identifying multiplex marker expression patterns from single EVs. For validation, the single EV heterogeneity of highly expressed cancer cell markers is profiled across different cancer cell lines. These findings exemplify squeezable MHPs as a robust platform with high sensitivity, multiplexing, and scalability for resolving single EV heterogeneity and advancing molecular assay technologies.
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Details
- Title
- Squeezable Hydrogel Microparticles for Single Extracellular Vesicle Protein Profiling
- Creators
- Yoon Ho Roh - University of PennsylvaniaRenee‐Tyler T. Morales - University of PennsylvaniaEmily Huynh - University of PennsylvaniaUday Chintapula - University of PennsylvaniaDavid E. Reynolds - University of PennsylvaniaRenis J. Agosto-Nieves - University of PennsylvaniaDaniel Oh - University of PennsylvaniaAkari J. Seiner - Drexel UniversityJianhua Lim - University of PennsylvaniaChristopher B. Rodell - Drexel UniversityJina Ko - University of Pennsylvania
- Publication Details
- Small (Weinheim an der Bergstrasse, Germany), e2407809
- Publisher
- WILEY-V C H VERLAG GMBH
- Number of pages
- 9
- Grant note
- PennPREP Scholarship: R00CA256353, DP2GM154018 NSF GRFP: R35GM147184, 23CDA1054367
Y.H.R. and R.T.T.M. contributed equally to this work. This work was supported by R00CA256353 and DP2GM154018 to YHR, RTM, EH, UC, DO, JL, and JK; NSF GRFP to DR; PennPREP Scholarship to RAN; a U.S. Department of Education pre-doctoral GAANN fellowship to AJS; R35GM147184 and 23CDA1054367 to CBR. The authors would like to thank Abhishek P. Dhand for discussing characterization of squeezable hydrogels and Luca Musante for discussing Nano-Flow Cytometry.
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- School of Biomedical Engineering, Science, and Health Systems
- Web of Science ID
- WOS:001343931900001
- Scopus ID
- 2-s2.0-85207324404
- Other Identifier
- 991021932215504721
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InCites Highlights
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Chemistry, Multidisciplinary
- Chemistry, Physical
- Materials Science, Multidisciplinary
- Nanoscience & Nanotechnology
- Physics, Applied
- Physics, Condensed Matter