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Updated: May 3, 2026
Isolation and Analysis of Plasma Lipoproteins by Ultracentrifugation
Published on: January 28, 2021
Proteomic analysis of proteins eliminated by low-density lipoprotein apheresis
Yumiko Yuasa1, Tsukasa Osaki, Hisashi Makino
1Department of Molecular Innovation in Lipidology, National Cerebral and Cardiovascular Center Research Institute, Osaka, Japan; Division of Nutrition and Metabolism, Department of Biophysics, Postgraduate School of Health Science, Kobe University, Kobe, Japan.
Insights
Low-density lipoprotein apheresis (LDL-A) removes more than LDL cholesterol. Analysis of LDL-A waste fluid identified novel proteins like vitronectin, potentially explaining its atherosclerosis-preventive effects.
Area of Science:
- Cardiovascular Medicine
- Proteomics
- Biochemistry
Background:
- Low-density lipoprotein apheresis (LDL-A) is established for managing familial hypercholesterolemia.
- Emerging evidence suggests LDL-A offers benefits beyond LDL cholesterol reduction.
- Atherosclerosis development involves complex molecular pathways influenced by various proteins.
Purpose of the Study:
- To investigate the comprehensive protein profile removed during LDL-A treatment.
- To identify novel proteins implicated in atherosclerosis pathogenesis that are targeted by LDL-A.
- To elucidate the mechanisms underlying LDL-A's therapeutic efficacy beyond LDL particle removal.
Main Methods:
- Proteomic analysis (2D electrophoresis and mass spectrometry) of LDL apheresis waste fluid.
- Quantification of specific protein levels (vitronectin, Apo C-III) via ELISA before and after LDL-A.
- Correlation analysis between protein removal and serum concentration changes.
Main Results:
- Identified 48 proteins in LDL-A waste fluid, including coagulation, inflammatory, and adhesion factors.
- Newly identified vitronectin and apolipoprotein C-III (Apo C-III) as proteins removed by LDL-A.
- Demonstrated significant reductions in serum vitronectin (82.4%) and Apo C-III (54.8%) post-treatment.
- Observed distinct removal patterns: Apo C-III with lipoproteins, vitronectin independently.
Conclusions:
- LDL-A treatment removes a broad spectrum of proteins beyond LDL particles.
- The removal of proteins like vitronectin and Apo C-III may contribute to LDL-A's atherosclerosis-preventive effects.
- Proteomic analysis of apheresis waste fluid offers insights into treatment mechanisms and efficacy.
Abstract:
Low-density lipoprotein apheresis (LDL-A) treatment has been shown to decrease serum LDL cholesterol levels and prevent cardiovascular events in homozygous patients with familial hypercholesterolemia. Recently, LDL-A treatment has been suggested to have beneficial effects beyond the removal of LDL particles. In this study, to clarify the preventive effects of LDL-A treatment on atherosclerosis, the waste fluid from the adsorption columns was analyzed. The waste fluid of LDL adsorption columns was analyzed by two-dimensional electrophoresis followed by mass spectrometry. Serum concentrations of the newly identified proteins before and after LDL-A treatment were measured by enzyme-linked immunosorbent assay. We identified 48 kinds of proteins in the waste fluid of LDL adsorption columns, including coagulation factors, thrombogenic factors, complement factors, inflammatory factors and adhesion molecules. In addition to the proteins that were reported to be removed by LDL-A treatment, we newly identified several proteins that have some significant roles in the development of atherosclerosis, including vitronectin and apolipoprotein C-III (Apo C-III). The serum levels of vitronectin and Apo C-III decreased by 82.4% and 54.8%, respectively, after a single LDL-A treatment. While Apo C-III was removed with very low-density lipoprotein (VLDL) and LDL, vitronectin was removed without association with lipoproteins. The removal of proteins observed in the waste fluid has a certain impact on their serum levels, and this may be related to the efficacy of LDL-A treatment. Proteomic analysis of the waste fluid of LDL adsorption columns may provide a rational means of assessing the effects of LDL-A treatment.
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