Related Experiment Video
Updated: Jan 11, 2026
06:47
Isolation and Analysis of Plasma Lipoproteins by Ultracentrifugation
Published on: January 28, 2021
12.3K
Simultaneous Isolation and Characterization of Lipoprotein Classes in Plasma, Including HDL Subclasses and the
Joanne Agus1, Olesia Gololobova2, Jack Zheng1
1Department of Nutrition, University of California Davis, California, United States of America.
Biorxiv : the Preprint Server for Biology
|November 19, 2025
Summary
Researchers developed a new method to isolate lipoprotein (LPP) classes from plasma, enabling detailed analysis of their RNA cargo for diagnostics and therapeutics. This technique successfully separates LPPs with minimal contamination.
Area of Science:
- Biochemistry
- Molecular Biology
- Nanotechnology
Background:
- Lipoproteins (LPPs) and extracellular vesicles transport extracellular small RNA, crucial for diagnostics and therapeutics.
- Isolating intact LPPs from plasma is challenging due to overlapping densities and sizes, hindering cargo analysis.
Purpose of the Study:
- To develop a method for simultaneous separation of LPP classes from plasma.
- To enable accurate characterization of LPP cargo for understanding their function.
Main Methods:
- Sequential flotation ultracentrifugation followed by size exclusion chromatography.
- Utilized western blot, gel electrophoresis, NMR, and electron microscopy for characterization.
- Method optimized for 500μL of starting plasma.
Main Results:
- Successfully separated LPP classes with minimal contamination.
- Demonstrated unique lipidomic, proteomic, and small RNA signatures for each LPP class.
- Identified very high-density high-density lipoprotein particles (1.21-1.25 g/mL) for the first time.
Conclusions:
- The developed method allows for efficient and clean isolation of LPP classes.
- Characterization of LPP subclasses reveals distinct molecular signatures.
- This advancement facilitates the study of LPPs in diagnostics and therapeutics.
