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Updated: Jan 11, 2026

Isolation and Analysis of Plasma Lipoproteins by Ultracentrifugation
Published on: January 28, 2021
Application of Polyacrylamide Gel Electrophoresis in Analyzing Lipoprotein Subfractions Relevant to Atherosclerotic
Ya-Nan Yao1, Ya-Long Liao1, Yun-Xiong Li1
1Laboratory Medicine, Guangdong Provincial People's Hospital (Guangdong Academy of Medical Sciences), Southern Medical University.
Insights
Small dense LDL (sdLDL) is a key driver of atherosclerotic cardiovascular disease (ASCVD). Polyacrylamide gel electrophoresis (PAGE) offers a practical method for detecting LDL subfractions to better assess cardiovascular risk.
Area of Science:
- Cardiovascular Science
- Biochemistry
- Medical Diagnostics
Background:
- Low-density lipoprotein cholesterol (LDL-C) is integral to atherosclerotic cardiovascular disease (ASCVD) development.
- LDL particle heterogeneity, specifically small dense LDL (sdLDL), significantly impacts atherosclerosis promotion.
- Elevated sdLDL, even with normal LDL-C, can lead to underestimation of cardiovascular risk in conditions like metabolic syndrome and diabetes.
Purpose of the Study:
- To highlight the clinical significance of LDL subfraction analysis for accurate ASCVD risk stratification.
- To introduce Polyacrylamide Gel Electrophoresis (PAGE) as a practical and effective method for LDL subfraction detection.
- To demonstrate the utility of PAGE in assessing cardiovascular risk based on LDL particle size.
Main Methods:
- Polyacrylamide Gel Electrophoresis (PAGE) utilizing non-denaturing gradient gels and Sudan black staining for lipoprotein separation.
- Analysis of lipoprotein particles based on molecular sieve effect and charge differences.
- Determination of mean LDL particle size from PAGE results.
Main Results:
- PAGE technology provides high-resolution and reproducible separation of LDL subfractions.
- Mean LDL particle sizes were determined for five samples, ranging from 275.3 Å to 252.9 Å.
- These particle sizes correlated with distinct cardiovascular disease risk levels: low, marginal, mild, moderate, and severe.
Conclusions:
- Accurate detection of LDL subfractions is crucial for precise cardiovascular risk assessment, especially in patients with dyslipidemia.
- PAGE technology presents a feasible and effective alternative to complex traditional methods for LDL subfraction analysis.
- PAGE-based lipoprotein typing can significantly aid clinicians in optimizing treatment strategies for ASCVD prevention.
Abstract:
Low-density lipoprotein cholesterol (LDL-C) is a key factor in the development of atherosclerotic cardiovascular disease (ASCVD). Elevated levels of LDL-C are closely linked to the formation of atherosclerotic plaques and the risk of cardiovascular events. However, the heterogeneity of LDL significantly influences its ability to promote atherosclerosis. Research indicates that Low-density lipoprotein (LDL) particles can be further categorized into small dense LDL (sdLDL) and large buoyant LDL (lbLDL). Among these, sdLDL is more likely to penetrate vascular endothelium and has stronger oxidative modification activity, which is particularly associated with ASCVD. In patients with metabolic syndrome, diabetes, and hypertriglyceridemia, LDL-C levels may be normal, but the proportion of sdLDL is elevated, leading to an underestimation of cardiovascular risk. Therefore, accurately detecting LDL subfractions is crucial for stratifying ASCVD risk and optimizing intervention strategies. Traditional LDL-C detection methods, such as the Friedewald formula and direct measurement, only reflect total cholesterol levels and cannot differentiate between subfractions. Methods for detecting LDL-C subfractions include ultracentrifugation (UC), high-performance liquid chromatography (HPLC), and nuclear magnetic resonance (NMR) techniques. However, these methods have high technical requirements, complex operations, or expensive equipment, making them impractical for widespread clinical use. Polyacrylamide gel electrophoresis (PAGE) technology separates lipoprotein particles through the molecular sieve effect and charge differences. This technique uses non-denaturing gradient gel systems, combined with specific stains like Sudan black, offering high-resolution, strong reproducibility, and ease of operation. Lipoprotein typing using PAGE technology can provide a more comprehensive and precise assessment of patients' lipoprotein levels, aiding in clinical treatment decisions. This article has presented five representative results. The mean LDL-particle size of sample A, B, C, D, E are 275.3 Å, 266.6 Å, 258.2 Å , 257.4 Å, and 252.9 Å, indicating a low, marginal, mild, moderate, and severe risk of cardiovascular disease due to dyslipidemia, respectively.
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