Comparative analysis of tangential flow filtration and ultracentrifugation, both combined with subsequent size exclusion chromatography, for the isolation of small extracellular vesicles

超离心机 大小排阻色谱法 纳米粒子跟踪分析 过滤(数学) 分析超速离心 色谱法 细胞外小泡 分离(微生物学) 化学 生物 生物化学 生物信息学 细胞生物学 微泡 数学 小RNA 统计 基因
作者
Kekoolani S Visan,Richard J. Lobb,Sunyoung Ham,Luize Gonçalves Lima,Carlos Palma,Chai Pei Zhi Edna,Li-Ying Wu,Harsha Gowda,Keshava K. Datta,Günter Härtel,Carlos Salomón,Andreas Möller
出处
期刊:Journal of extracellular vesicles [Wiley]
卷期号:11 (9) 被引量:25
标识
DOI:10.1002/jev2.12266
摘要

Abstract Small extracellular vesicles (sEVs) provide major promise for advances in cancer diagnostics, prognostics, and therapeutics, ascribed to their distinctive cargo reflective of pathophysiological status, active involvement in intercellular communication, as well as their ubiquity and stability in bodily fluids. As a result, the field of sEV research has expanded exponentially. Nevertheless, there is a lack of standardisation in methods for sEV isolation from cells grown in serum‐containing media. The majority of researchers use serum‐containing media for sEV harvest and employ ultracentrifugation as the primary isolation method. Ultracentrifugation is inefficient as it is devoid of the capacity to isolate high sEV yields without contamination of non‐sEV materials or disruption of sEV integrity. We comprehensively evaluated a protocol using tangential flow filtration and size exclusion chromatography to isolate sEVs from a variety of human and murine cancer cell lines, including HeLa, MDA‐MB‐231, EO771 and B16F10. We directly compared the performance of traditional ultracentrifugation and tangential flow filtration methods, that had undergone further purification by size exclusion chromatography, in their capacity to separate sEVs, and rigorously characterised sEV properties using multiple quantification devices, protein analyses and both image and nano‐flow cytometry. Ultracentrifugation and tangential flow filtration both enrich consistent sEV populations, with similar size distributions of particles ranging up to 200 nm. However, tangential flow filtration exceeds ultracentrifugation in isolating significantly higher yields of sEVs, making it more suitable for large‐scale research applications. Our results demonstrate that tangential flow filtration is a reliable and robust sEV isolation approach that surpasses ultracentrifugation in yield, reproducibility, time, costs and scalability. These advantages allow for implementation in comprehensive research applications and downstream investigations.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
大幅提高文件上传限制,最高150M (2024-4-1)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
刚刚
yyds完成签到,获得积分10
1秒前
1秒前
怕黑网络关注了科研通微信公众号
2秒前
小二郎应助迷你的豪英采纳,获得10
2秒前
secret发布了新的文献求助10
2秒前
WzH发布了新的文献求助10
2秒前
刘济源发布了新的文献求助20
2秒前
耍酷依玉完成签到,获得积分20
3秒前
冲冲冲完成签到 ,获得积分10
3秒前
sjx发布了新的文献求助10
3秒前
3秒前
小胡同学给小胡同学的求助进行了留言
3秒前
沈嘀嘀完成签到,获得积分10
3秒前
4秒前
丘比特应助爱学习的小白采纳,获得10
4秒前
yile完成签到,获得积分10
4秒前
有魅力荟完成签到,获得积分10
5秒前
5秒前
耍酷依玉发布了新的文献求助10
5秒前
6秒前
甜甜玫瑰发布了新的文献求助10
6秒前
finger完成签到,获得积分10
7秒前
曾梦发布了新的文献求助10
7秒前
7秒前
8秒前
8秒前
三石完成签到,获得积分10
9秒前
9秒前
风中虔纹完成签到,获得积分10
9秒前
罗预言本罗完成签到,获得积分10
10秒前
小牙医发布了新的文献求助10
10秒前
nkmenghan发布了新的文献求助10
10秒前
刚子发布了新的文献求助10
11秒前
春鸮鸟完成签到 ,获得积分10
11秒前
潇潇雨落发布了新的文献求助10
11秒前
11秒前
zhuyinghao发布了新的文献求助50
11秒前
12秒前
鹿死发布了新的文献求助10
12秒前
高分求助中
The late Devonian Standard Conodont Zonation 2000
The Lali Section: An Excellent Reference Section for Upper - Devonian in South China 1500
Nickel superalloy market size, share, growth, trends, and forecast 2023-2030 1000
Smart but Scattered: The Revolutionary Executive Skills Approach to Helping Kids Reach Their Potential (第二版) 1000
Mantiden: Faszinierende Lauerjäger Faszinierende Lauerjäger 800
PraxisRatgeber: Mantiden: Faszinierende Lauerjäger 800
A new species of Coccus (Homoptera: Coccoidea) from Malawi 500
热门求助领域 (近24小时)
化学 医学 生物 材料科学 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 基因 遗传学 催化作用 物理化学 免疫学 量子力学 细胞生物学
热门帖子
关注 科研通微信公众号,转发送积分 3245070
求助须知:如何正确求助?哪些是违规求助? 2888780
关于积分的说明 8255477
捐赠科研通 2557124
什么是DOI,文献DOI怎么找? 1385882
科研通“疑难数据库(出版商)”最低求助积分说明 650248
邀请新用户注册赠送积分活动 626457