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Related Concept Videos

Overview of Secretory Vesicles01:33

Overview of Secretory Vesicles

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Secretory vesicles, also known as dense core vesicles (DCVs), are membrane-bound vesicles that transport secretory proteins, such as hormones or neurotransmitters. Regulated secretory vesicles transport proteins from the trans-Golgi network to the exterior of the cell. Proteins present in regulated secretory vesicles are required to be rapidly exocytosed in large amounts upon a specific stimulus.
Various proteins regulate the aggregation of molecules inside the secretory vesicles. Chromogranins...
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Scalable Biomanufacturing Workflow to Produce and Isolate Natural Killer Cell-Derived Extracellular Vesicle-Based Cancer Biotherapeutics
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Production of Genetically Engineered Extracellular Vesicles for Targeted Protein Delivery.

Leyla A Ovchinnikova1, Evgeniy G Evtushenko2, Dmitriy V Bagrov3

  • 1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, Russia.

Bio-Protocol
|November 13, 2025
PubMed
Summary
This summary is machine-generated.

Genetically engineered extracellular vesicles (EVs) can now be precisely targeted to specific cells, like antigen-presenting cells (APCs), using a new modular system for enhanced protein delivery. This method ensures reliable production and characterization of therapeutic EVs.

Keywords:
Antigen-presenting cellsCD206EVsEngineered EVsNTAProtein deliveryTarget deliveryVSV-G

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Nanomedicine

Background:

  • Extracellular vesicles (EVs) are natural carriers for intercellular communication and show potential for targeted drug delivery.
  • Previous work established the feasibility of using genetically engineered EVs for therapeutic protein delivery.
  • Developing methods for specific cell targeting and robust production of engineered EVs is crucial for clinical translation.

Purpose of the Study:

  • To detail a protocol for generating mannose receptor (CD206)-targeted extracellular vesicles (EVs) using a modular plasmid system.
  • To enable customizable EV surface modification for targeting specific cell types, such as antigen-presenting cells (APCs).
  • To provide a flexible and reproducible platform for engineering, isolating, and characterizing therapeutic EVs.

Main Methods:

  • Utilized a three-plasmid modular system for customizable EV budding, cargo loading, and CD206-specific surface targeting in HEK293T cells.
  • Employed differential centrifugation and chromatography for EV isolation.
  • Characterized EVs using transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA), and validated uptake in primary human dendritic cells.

Main Results:

  • Successfully generated genetically engineered EVs displaying CD206-targeting moieties.
  • Established a robust workflow for EV isolation and multi-modal characterization, ensuring reproducibility.
  • Demonstrated functional uptake of engineered EVs by primary human activated dendritic cells.

Conclusions:

  • This protocol provides a detailed and flexible method for producing targeted EVs for therapeutic applications.
  • The modular platform allows for easy adaptation to different target cells and therapeutic cargoes.
  • The developed system integrates reliable EV engineering with gold-standard isolation and characterization techniques, paving the way for advanced nanomedicine strategies.