Abstract
High-density lipoprotein (HDL) and exosomes are promising sources of biomarkers. However, the limited sample volume and access to the ultracentrifuge equipment are still an issue during HDL and exosome isolation. This study aimed to isolate HDL and exosomes using an ultracentrifugation-free method with various small serum volumes. HDL was isolated from 200 µL (HDL200) and 500 µL (HDL500) of sera. Three different volumes: 50 µL (Exo50), 100 µL (Exo100), and 250 µL (Exo250) were used for exosome isolation. HDL and exosomes were isolated using commercial kits with the modified method and characterized by multiple approaches. The HDL levels of HDL200 and HDL500 were not significantly different (p > 0.05), with percent recoveries of >90%. HDL200 and HDL500 had the same protein pattern with a biochemical similarity of 99.60 ± 0.10%. The particle sizes of Exo50, Exo100, and Exo250 were in the expected range. All isolated exosomes exhibited a similar protein pattern with a biochemical similarity of >99%. In conclusion, two different serum volumes (200 and 500 µL) and three different serum volumes (50, 100, and 250 µL) can be employed for HDL and exosome isolation, respectively. The possibility of HDL and exosome isolation with small volumes will accelerate biomarker discoveries with various molecular diagnostic approaches.
Highlights
The essential key player of the molecular diagnosis of diseases is mainly supported by a variety of biomarkers [1]
The use of proteomic studies on High-density lipoprotein (HDL) particles has identified 85 proteins associated with lipid metabolism and transport, hemostasis, immune response, metal binding, and vitamin transport [6]
We demonstrated an ultracentrifugation-free method to isolate the HDL and exosome from serum samples with a variety of small volumes, as well as their characterizations
Summary
The essential key player of the molecular diagnosis of diseases is mainly supported by a variety of biomarkers [1]. Body fluids are still a significant primary source of biomarkers for noninvasive clinical diagnostic needs [2,3]. Body fluid-derived soluble biomarkers possess much information, as they provide significant evidence related to the health status [4]. High-density lipoprotein (HDL) particles are complex molecules that transport lipids and other components (proteins, hormones, and vitamins, as well as microRNAs (miRNAs)) to target tissues and cells [5]. Vickers et al (2011) discovered that the HDL miRNA profile of healthy people was significantly different from that of people with familial hypercholesterolemia [9]. Wagner et al (2013) observed that the absolute copy numbers of miR-30c, miR-92a, and miR-146 in HDL isolated from patients with acute coronary syndrome were significantly different from those in healthy people [7]. Changes in HDL components are clearly related to the health status
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