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In vitro and in cellulo ApoE particle formation, isolation, and characterization
Karina Lindner1, Larissa C van Ek1, Khader Awwad2
1Department of Cell Physiology and Metabolism, Faculty of Medicine, University of Geneva, Rue Michel-Servet 1, 1211 Geneva, Switzerland.
STAR Protocols
|January 3, 2023
Summary
This study outlines methods to analyze Apolipoprotein E (ApoE) particles, crucial for lipid transport. Researchers detail techniques for assessing both in vitro and in cellulo ApoE particles and their lipid cargo.
Area of Science:
- Biochemistry
- Cell Biology
- Lipid Metabolism
Background:
- Apolipoprotein E (ApoE) is essential for lipid transport in aqueous environments.
- Understanding ApoE particle formation and lipid cargo is vital for metabolic research.
Purpose of the Study:
- To provide detailed protocols for assessing in vitro and in cellulo Apolipoprotein E (ApoE) particles.
- To describe methods for analyzing the lipid cargo of ApoE particles.
Main Methods:
- In vitro assessment of ApoE particles using right-angle and dynamic light scattering.
- Generation of in cellulo ApoE particles under fatty-acid-induced stress.
- Isolation of ApoE particles via immunoprecipitation from cell secretome.
- Analysis of lipid cargo using thin-layer chromatography.
Main Results:
- Established protocols for characterizing ApoE particle formation and size.
- Demonstrated methods for generating and isolating cellular ApoE particles.
- Enabled detailed analysis of lipid cargo within ApoE particles.
Conclusions:
- The described methods facilitate comprehensive study of Apolipoprotein E particle assembly and lipid transport.
- These protocols can advance research into lipid-related diseases and metabolic disorders.
