Stimuli‐Mediated Specific Isolation of Exosomes from Blood Plasma for High‐Throughput Profiling of Cancer Biomarkers

Abstract

Exosomes, ranging from 30–150 nm in diameter, have emerged as promising non‐invasive biomarkers for the diagnosis and prognosis of numerous diseases. However, current research on exosomes is largely restricted by the lack of an efficient method to isolate exosomes from real samples. Herein, the first stimuli‐mediated enrichment and purification system to selectively and efficiently extract exosomes from clinical plasma for high‐throughput profiling of exosomal mRNAs as cancer biomarkers is presented. This novel isolation system relies on specific installation of the stimuli‐responsive copolymers onto exosomal phospholipid bilayers, by which the enrichment and purification are exclusively achieved for exosomes rather than the non‐vesicle counterparts co‐existing in real samples. The stimuli‐mediated isolation system outperforms conventional methods such as ultracentrifugation and polyethylene glycol‐based precipitation in terms of isolation yield, purity, and retained bioactivity. The high performance of the isolation system is demonstrated by enriching exosomes from 77 blood plasma samples and validated the clinical potentials in profiling exosomal mRNAs for cancer diagnosis and discrimination with high accuracy. This simple isolation system can boost the development of extracellular vesicle research, not limited to exosomes, in both basic and clinical settings.

Identification of faecal extracellular vesicles as novel biomarkers for the non‐invasive diagnosis and prognosis of colorectal cancer

“…Apart from EV protein, RNA is another vital aspect of EV candidate biomarkers. With the assistance of RNA sequencing (RNA‐Seq) and quantitative reverse transcription PCR (RT‐qPCR), RNA biomarkers are of increasing interest and are in rapid development (He et al., 2020; Liu et al., 2022; Min et al., 2019; Ostenfeld et al., 2016; Wang et al., 2022). Although faecal RNAs have been revealed as biomarkers for CRC (Duran‐Sanchon et al., 2020; Herring et al., 2021; Wu et al., 2012; Yau et al., 2014), few have concentrated on RNAs in isolated fEVs.…”

“…EVs inherit the specific molecular information from their parental cells and are thus regarded as promising cancer biomarkers for “liquid biopsy” (Shah et al., 2018; Yu et al., 2022; Zhou et al., 2020). Plasma EVs have been broadly applied for cancer diagnosis and prognosis (Liu et al., 2022; Park et al., 2021; Whiteside, 2020). Since blood circulates throughout the body, plasma EVs are derived from diverse cells and tissues (Alberro et al., 2021).…”

Extracellular vesicle isolation, purification and evaluation in cancer diagnosis

“…Development of such systems will definitely boost research regarding the EVs in basic and clinics, and can be a substitute for conventional methods, such as precipitation and UC in terms of purity, isolation yield and retained bioactivity (Ref. 137). Another advance in the field is the use of reference biological materials, such as recombinant EV (rEV), which has biophysical and biochemical traits of sample EVs.…”

Reversible zwitterionic coordination enables rapid, high-yield, and high-purity isolation of extracellular vesicles from biofluids

“…Tumor-derived EVs have been explored as potential biomarkers for liquid biopsies in cancer patients ( 16 ). These EVs have been frequently found in cancer patient’s serum (such as breast cancer and liver cancer) ( 57 , 58 ), urine (such as kidney cancer and bladder cancer) ( 28 ), and saliva (such as nasopharynx cancer and oral cancer) ( 7 ). Considering the low concentration of EVs and the interference of a large number of soluble proteins in body fluids, it is necessary to develop a high-yield and high-purity isolation method for extracting EVs from different body fluids for biomarker identification and cancer diagnostics.…”