Integrated microfluidic system for isolating exosome and analyzing protein marker PD-L1

Lu, Yunxing; Ye, Ling; Jian, Xiaoyu; Yang, Dawei; Zhang, Hongwei; Tong, Zhaoduo; Wu, Zhenhua; Shi, Nan; Han, Yunwei; Mao, Hongju

doi:10.1016/j.bios.2021.113879

Cited by 37 publications

(34 citation statements)

References 50 publications

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“…To reveal the fundamental characteristics and functions of PD-L1-positive sEVs, it is necessary to separate and obtain them from the heterogeneous sEV populations. Currently, immunoaffinity-based isolation techniques are still the primary strategies for isolating sEV subpopulations including PD-L1-positive sEVs, mostly for subsequent quantitative detection. , However, the sEVs are trapped by isolation carriers and PD-L1 is blocked by antibody, which seriously limits further functional research. Traceless release after capture of PD-L1-positive sEVs is critical for the following in-depth analyses, especially for unmasking their biological behaviors and functions.…”

Section: Resultsmentioning

confidence: 99%

Aptamer-Assisted Traceless Isolation of PD-L1-Positive Small Extracellular Vesicles for Dissecting Their Subpopulation Signature and Function

Liu

et al. 2022

Anal. Chem.

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Small extracellular vesicles (sEVs) are heterogeneous membrane-bound vesicles that carry numerous bioactive molecules. Studies have reported that sEVs carrying PD-L1 on the surface could contribute to immunosuppression; however, the precise mechanisms are unclear. To fully dissect their mode of action, it requires qualified methods to specifically isolate natural PD-L1-positive sEVs from heterogeneous sEVs. This study reported an aptamer-assisted capture-and-release strategy for traceless isolation of PD-L1-positive sEVs. The PD-L1 aptamer-anchored magnetic microspheres enable the specific capture of PD-L1-positive sEVs. The traceless release of captured PD-L1-positive sEVs was triggered by competition of complementary oligonucleotides, endowing the obtained label-free PD-L1-positive sEVs with natural properties. Benefited from this traceless isolation strategy, the distinct molecule profiles in adhesion and immuno-regulation between PD-L1-positive and PD-L1-negative sEVs were revealed. Compared to PD-L1-negative sEVs, PD-L1-positive sEVs were much more concentrated in cadherin binding, accompanied by increased adhesion to lymphatic endothelial cells and T cells but decreased adhesion to the extracellular matrix. Moreover, PD-L1-positive sEVs could transfer their enriched immunosuppressive “synapse”-related proteins to antigen-presenting cells, thereby inducing a tolerogenic-like phenotype. In summary, the present work dissects the subpopulation signature and action mode of PD-L1-positive sEVs for the first time and provides a general approach to the traceless isolation of sEV subpopulations.

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Section: Resultsmentioning

confidence: 99%

Aptamer-Assisted Traceless Isolation of PD-L1-Positive Small Extracellular Vesicles for Dissecting Their Subpopulation Signature and Function

Liu

et al. 2022

Anal. Chem.

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“…The practice of exosomes in clinical applications (such as point-of-care testing, POCT) is seriously hindered. In recent years, with the development of microfluidic chips (Lu et al, 2022), it has become possible to handle or manipulate small volumes of liquid in microplates and microchannels. Microfluidic devices are used to achieve the integration of isolation and identification on a microchip (Ullah Khan et al, 2021).…”

Section: Emerging Methods For Exosome Isolationmentioning

confidence: 99%

Recent developments in isolating methods for exosomes

Gao

et al. 2023

Front. Bioeng. Biotechnol.

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Exosomes are the smallest extracellular vesicles that can be released by practically all cell types, and range in size from 30 nm to 150 nm. As the major marker of liquid biopsies, exosomes have great potential for disease diagnosis, therapy, and prognosis. However, their inherent heterogeneity, the complexity of biological fluids, and the presence of nanoscale contaminants make the isolation of exosomes a great challenge. Traditional isolation methods of exosomes are cumbersome and challenging with complex and time-consuming operations. In recent years, the emergence of microfluidic chips, nanolithography, electro-deposition, and other technologies has promoted the combination and innovation of the isolation methods. The application of these methods has brought very considerable benefits to the isolation of exosomes such as ultra-fast, portable integration, and low loss. There are significant functional improvements in isolation yield, isolation purity, and clinical applications. In this review, a series of methods for the isolation of exosomes are summarized, with emphasis on the emerging methods, and in-depth comparison and analysis of each method are provided, including their principles, merits, and demerits.

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“…One of the significant advantages of traditional channel-based microfluidic chips for LOC applications is the use of patterned microstructures or modifications as processing units in the flow channel. Such structures are critical for mixing, 116 sample isolation & enrichment, 117 capture, 118 detection, 119 and other purposes at the microscale. As an active actuation approach, digital microfluidics, however, no longer need microstructures for basic fluidic manipulation.…”

Section: Chip-scale Integrationmentioning

confidence: 99%

Combining sensors and actuators with electrowetting-on-dielectric (EWOD): advanced digital microfluidic systems for biomedical applications

Tong

Shen

et al. 2023

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Integrated microfluidic system for isolating exosome and analyzing protein marker PD-L1

Cited by 37 publications

References 50 publications

Aptamer-Assisted Traceless Isolation of PD-L1-Positive Small Extracellular Vesicles for Dissecting Their Subpopulation Signature and Function

Aptamer-Assisted Traceless Isolation of PD-L1-Positive Small Extracellular Vesicles for Dissecting Their Subpopulation Signature and Function

Recent developments in isolating methods for exosomes

Combining sensors and actuators with electrowetting-on-dielectric (EWOD): advanced digital microfluidic systems for biomedical applications

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