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Quantifying the Endosomal Escape of pH-Responsive Nanoparticles Using the Split Luciferase Endosomal Escape
Maximilian A Beach1, Serena L Y Teo2, Moore Z Chen2
1Department of Chemistry, The University of Melbourne, Parkville, Victoria 3010, Australia.
ACS Applied Materials & Interfaces
|December 29, 2021
Summary
Developing new nanoparticles for drug delivery is crucial. A novel assay, split luciferase endosomal escape quantification (SLEEQ), directly measures how effectively nanoparticles release therapeutic cargo into cells, improving drug delivery vehicle design.
Area of Science:
- Biotechnology
- Nanomedicine
- Drug Delivery
Background:
- Nanoparticles offer revolutionary therapeutic cargo delivery (peptides, proteins, RNA).
- Effective cytosolic delivery from nanoparticles remains a significant challenge for efficient drug vehicles.
- Current endosomal escape assays lack quantitative data, hindering nanoparticle design.
Purpose of the Study:
- To adapt the split luciferase endosomal escape quantification (SLEEQ) assay for direct, quantitative measurement of nanoparticle-induced endosomal escape.
- To evaluate the endosomal escape behavior of pH-responsive nanoparticles using the adapted SLEEQ assay.
Main Methods:
- Adapted the split luciferase endosomal escape quantification (SLEEQ) assay for direct measurement of cytosolic delivery.
- Applied SLEEQ to assess two pH-responsive nanoparticles: poly(2-diisopropylamino ethyl methacrylate) (PDPAEMA) core and a 1:1 ratio of poly(2-diethylamino ethyl methacrylate) (PDEAEMA) and PDPAEMA.
Main Results:
- The adapted SLEEQ assay directly quantified cytosolic delivery of cargo from nanoparticles.
- Engineering the nanoparticle disassembly pH improved endosomal escape efficiency by fivefold.
- Demonstrated the utility of SLEEQ for evaluating nanoparticle-based drug delivery systems.
Conclusions:
- The SLEEQ assay provides a versatile and sensitive method for directly measuring nanoparticle-mediated endosomal escape.
- Optimizing nanoparticle disassembly pH is a key strategy for enhancing endosomal escape efficiency.
- This advancement will accelerate the development of more effective nanoparticle drug delivery vehicles.
