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Related Experiment Video

Updated: Aug 5, 2025

Obtention of Giant Unilamellar Hybrid Vesicles by Electroformation and Measurement of their Mechanical Properties by Micropipette Aspiration
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Measuring Encapsulation Efficiency in Cell-Mimicking Giant Unilamellar Vesicles.

Pashiini Supramaniam1, Zibo Wang2,3, Stelios Chatzimichail2

  • 1Department of Chemistry, Imperial College London, London W12 0BZ, U.K.

ACS Synthetic Biology
|March 28, 2023
PubMed
Summary
This summary is machine-generated.

We developed a new method to count molecules inside artificial cells. This quality control protocol precisely measures encapsulation efficiency per vesicle, crucial for synthetic biology applications.

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Area of Science:

  • Synthetic biology
  • Biotechnology
  • Biophysics

Background:

  • Bottom-up synthetic biology aims to create programmable artificial cells.
  • Existing methods for generating giant unilamellar vesicle-based artificial cells are numerous.
  • Quantitative measurement of molecular constituents in newly formed artificial cells remains underdeveloped.

Purpose of the Study:

  • To develop and validate an artificial cell quality control (AC/QC) protocol.
  • To enable absolute quantification of encapsulated biomolecules in single vesicles.
  • To assess the variability of encapsulation efficiency in artificial cells.

Main Methods:

  • Utilized a microfluidic-based single-molecule approach for analysis.
  • Developed an artificial cell quality control (AC/QC) protocol.
  • Quantified encapsulated biomolecules on a per-vesicle basis.

Main Results:

  • Achieved absolute quantification of encapsulated biomolecules.
  • Determined per-vesicle encapsulation efficiencies, revealing significant variability (2.4% to 41%).
  • Demonstrated that desired internal concentrations can be achieved by adjusting initial concentrations, despite variability.

Conclusions:

  • The developed AC/QC protocol provides precise measurement of artificial cell contents.
  • Significant variability in encapsulation efficiency necessitates caution when using artificial cells as models or standards.
  • This method is vital for advancing the development of functional artificial cells in synthetic biology.