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Article

Design and Evaluation of Engineered Extracellular Vesicle (EV)-Based Targeting for EGFR-Overexpressing Tumor Cells Using Monobody Display

1
Institute for Quantitative Health Science and Engineering (IQ), Michigan State University, East Lansing, MI 48824, USA
2
Department of Biomedical Engineering, Michigan State University, East Lansing, MI 48824, USA
3
Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI 48824, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Jarek Baran
Bioengineering 2022, 9(2), 56; https://doi.org/10.3390/bioengineering9020056
Received: 20 December 2021 / Revised: 13 January 2022 / Accepted: 15 January 2022 / Published: 29 January 2022
Background: Extracellular vesicles (EVs) are attracting interest as a new class of drug delivery vehicles due to their intrinsic nature of biomolecular transport in the body. We previously demonstrated that EV surface modification with tissue-specific molecules accomplished targeted EV-mediated DNA delivery. Methods: Here, we describe reliable methods for (i) generating EGFR tumor-targeting EVs via the display of high-affinity monobodies and (ii) in vitro measurement of EV binding using fluorescence and bioluminescence labeling. Monobodies are a well-suited class of small (10 kDa) non-antibody scaffolds derived from the human fibronectin type III (FN3) domain. Results: The recombinant protein consists of the EGFR-targeting monobody fused to the EV-binding domain of lactadherin (C1C2), enabling the monobody displayed on the surface of the EVs. In addition, the use of bioluminescence or fluorescence molecules on the EV surface allows for the assessment of EV binding to the target cells. Conclusions: In this paper, we describe methods of EV engineering to generate targeted delivery vehicles using monobodies that will have diverse applications to furnish future EV therapeutic development, including qualitative and quantitative in vitro evaluation for their binding capacity. View Full-Text
Keywords: extracellular vesicles; EV engineering; EGFR targeting extracellular vesicles; EV engineering; EGFR targeting
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MDPI and ACS Style

Komuro, H.; Aminova, S.; Lauro, K.; Woldring, D.; Harada, M. Design and Evaluation of Engineered Extracellular Vesicle (EV)-Based Targeting for EGFR-Overexpressing Tumor Cells Using Monobody Display. Bioengineering 2022, 9, 56. https://doi.org/10.3390/bioengineering9020056

AMA Style

Komuro H, Aminova S, Lauro K, Woldring D, Harada M. Design and Evaluation of Engineered Extracellular Vesicle (EV)-Based Targeting for EGFR-Overexpressing Tumor Cells Using Monobody Display. Bioengineering. 2022; 9(2):56. https://doi.org/10.3390/bioengineering9020056

Chicago/Turabian Style

Komuro, Hiroaki, Shakhlo Aminova, Katherine Lauro, Daniel Woldring, and Masako Harada. 2022. "Design and Evaluation of Engineered Extracellular Vesicle (EV)-Based Targeting for EGFR-Overexpressing Tumor Cells Using Monobody Display" Bioengineering 9, no. 2: 56. https://doi.org/10.3390/bioengineering9020056

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