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Real-Time Quantification of Cell Internalization Kinetics by Bioluminescent Probes
Roxanne Castillo1, Di Wu1, Zheng Cao1
1Department of Chemical and Biomolecular Engineering, School of Engineering, University of California, Los Angeles, Los Angeles, CA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|July 14, 2022
Summary
This study introduces a novel method to track cellular delivery in real-time using bioluminescent nano-encapsulated enzymes. This approach quantifies internalization kinetics, advancing therapeutic vector development.
Area of Science:
- Biotechnology
- Cell Biology
- Biochemistry
Background:
- Effective intracellular delivery of therapeutic agents is crucial for various biomedical applications.
- Current methods for evaluating cellular uptake lack real-time, quantitative insights.
- Understanding cellular internalization dynamics is key to improving drug and gene delivery systems.
Purpose of the Study:
- To develop and validate a real-time method for quantifying cellular internalization kinetics.
- To utilize nano-encapsulated bioluminescent enzymes as probes for dynamic cellular uptake studies.
- To provide a quantitative framework for assessing intracellular delivery efficiency.
Main Methods:
- Synthesis and characterization of nano-encapsulated Firefly Luciferase (FLuc) enzymes.
- Real-time monitoring of bioluminescence (BL) in cells following delivery.
- Analysis of BL profiles to extract key kinetic parameters of cell internalization.
Main Results:
- Demonstrated successful encapsulation and cellular delivery of FLuc enzymes.
- Established a protocol for real-time BL assays to monitor internalization.
- Quantified cell internalization kinetics, providing dynamic measures of uptake.
Conclusions:
- Real-time quantification of internalization kinetics offers a significant advancement over conventional methods.
- This approach provides mechanistic insights into membrane translocation and cellular metabolism.
- The method is critical for the clinical development and optimization of effective delivery vectors.
Keywords:
Bioluminescence (BL)Cell internalization kineticsFirefly luciferase (FLuc)NanocapsulesReal-time quantification
