Co-isolation of extracellular vesicles and high-density lipoproteins using density gradient ultracentrifugation
نویسندگان
چکیده
Extracellular vesicles (EVs) facilitate intercellular communication by carrying bioactive molecules such as proteins, messenger RNA, and micro (mi)RNAs. Recently, high-density lipoproteins (HDL) isolated from human plasma were also reported to transport miRNA to other cells. HDL, when isolated from human plasma, ranges in density between 1.063 and 1.21 g/mL, which grossly overlap with the reported density of EVs. Consequently, HDL and EV will be co-isolated when using density gradient ultracentrifugation. Thus, more stringent isolation/separation procedures of EV and HDL are essential to know their relative contribution to the pool of circulating bioactive molecules.
منابع مشابه
Single-step isolation of extracellular vesicles by size-exclusion chromatography
BACKGROUND Isolation of extracellular vesicles from plasma is a challenge due to the presence of proteins and lipoproteins. Isolation of vesicles using differential centrifugation or density-gradient ultracentrifugation results in co-isolation of contaminants such as protein aggregates and incomplete separation of vesicles from lipoproteins, respectively. AIM To develop a single-step protocol...
متن کاملThe impact of disparate isolation methods for extracellular vesicles on downstream RNA profiling
Despite an enormous interest in the role of extracellular vesicles, including exosomes, in cancer and their use as biomarkers for diagnosis, prognosis, drug response and recurrence, there is no consensus on dependable isolation protocols. We provide a comparative evaluation of 4 exosome isolation protocols for their usability, yield and purity, and their impact on downstream omics approaches fo...
متن کاملUltracentrifugation in swinging-bucket and fixed-angle rotors evaluated for isolation and determination of high-density lipoprotein subfractions HDL2 and HDL3.
We evaluated the density-gradient ultracentrifugation method in a swinging-bucket rotor (Anal Biochem 111, 149-157, 1981) in a slightly modified version for isolation and determination of high-density lipoprotein (HDL) subfractions. We prestained the serum with Coomassie Brilliant Blue R, which did not change the hydrated densities of the lipoproteins, and after only 2.2 X 10(8) gav . min obtai...
متن کاملRapid preparative isolation of concentrated low density lipoproteins and of lipoprotein-deficient serum using vertical rotor gradient ultracentrifugation.
In order to study cellular metabolism of low density lipoproteins (LDL), ultracentrifugal methods have been used to isolate the lipoproteins. The use of vertical rotor ultracentrifugation very quickly produces small quantities of diluted lipoproteins per gradient, as well as small volumes of lipoprotein-deficient serum. We present modifications to this method in order to prepare routinely more ...
متن کاملMethod for lipoprotein(a) density profiling by BiEDTA differential density lipoprotein ultracentrifugation.
In this article, we demonstrate the analytical power of linking density gradient ultracentrifugation with affinity separations. Here we address some of the analytical challenges in the study of lipoprotein(a), (Lp(a)). The mean density distribution of Lp(a) was determined by a differential density lipoprotein profile (DDLP). For DDLP, the lipoprotein density distribution of a serum sample with ...
متن کامل