Extracellular vesicles (EVs) play an important role as active messengers in intercellular communication and distant microenvironment modeling. Increasingly, these EVs are recognized as important biomarkers for clinical diagnostics. However, current isolation methods of EVs are time-consuming and ineffective due to the high diffusive characteristics of nanoparticles coupled with fluid flow instability. Here, we develop a microfluidic CEntrifugal Nanoparticles Separation and Extraction (µCENSE) platform for the rapid and label-free isolation of microvesicles. By utilizing centrifugal microhydrodynamics, we subject the nanosuspensions between 100 nm and 1000 nm to a unique fluid flow resulting in a zonal separation into different outlets for easy post-processing. Our centrifugal platform utilizes a gentle and efficient size-based separation without the requirements of syringe pump and other accessories. Based on our results, we report a high separation efficiency of 90% and an extraction purity of 85% within a single platform. Importantly, we demonstrate high EV extraction using a table top centrifuge within a short duration of eight minutes. The simple processes and the small volume requirement further enhance the utility of the platform. With this platform, it serves as a potential for liquid biopsy extraction and point-of-care diagnostics.

1.
C.
Braicu
,
C.
Tomuleasa
,
P.
Monroig
,
A.
Cucuianu
,
I.
Berindan-Neagoe
, and
G. A.
Calin
,
Cell Death Differ.
22
(
1
),
34
45
(
2015
).
2.
D. D.
Taylor
and
C.
Gercel-Taylor
,
Front. Genet.
4
,
142
(
2013
).
3.
E.
Cocucci
,
G.
Racchetti
, and
J.
Meldolesi
,
Trends Cell Biol.
19
(
2
),
43
51
(
2009
).
4.
G.
Raposo
and
W.
Stoorvogel
,
J. Cell Biol.
200
(
4
),
373
(
2013
).
5.
H.
Valadi
,
K.
Ekström
,
A.
Bossios
,
M.
Sjöstrand
,
J. J.
Lee
, and
J. O.
Lötvall
,
Nat. Cell Biol.
9
(
6
),
654
659
(
2007
).
6.
J.
Lötvall
,
A. F.
Hill
,
F.
Hochberg
,
E. I.
Buzás
,
D.
Di Vizio
,
C.
Gardiner
,
Y. S.
Gho
,
I. V.
Kurochkin
,
S.
Mathivanan
,
P.
Quesenberry
,
S.
Sahoo
,
H.
Tahara
,
M. H.
Wauben
,
K. W.
Witwer
, and
C.
Théry
,
J. Extracell. Vesicles
3
(
1
),
26913
(
2014
).
7.
N.
Kosaka
,
H.
Iguchi
, and
T.
Ochiya
,
Cancer Sci.
101
(
10
),
2087
2092
(
2010
).
8.
B. K.
Thakur
,
H.
Zhang
,
A.
Becker
,
I.
Matei
,
Y.
Huang
,
B.
Costa-Silva
,
Y.
Zheng
,
A.
Hoshino
,
H.
Brazier
,
J.
Xiang
,
C.
Williams
,
R.
Rodriguez-Barrueco
,
J. M.
Silva
,
W.
Zhang
,
S.
Hearn
,
O.
Elemento
,
N.
Paknejad
,
K.
Manova-Todorova
,
K.
Welte
,
J.
Bromberg
,
H.
Peinado
, and
D.
Lyden
,
Cell Res.
24
(
6
),
766
769
(
2014
).
9.
H.
Shao
,
J.
Chung
,
L.
Balaj
,
A.
Charest
,
D. D.
Bigner
,
B. S.
Carter
,
F. H.
Hochberg
,
X. O.
Breakefield
,
R.
Weissleder
, and
H.
Lee
,
Nat. Med.
18
(
12
),
1835
1840
(
2012
).
10.
R.
Xu
,
D. W.
Greening
,
A.
Rai
,
H.
Ji
, and
R. J.
Simpson
,
Methods
87
,
11
25
(
2015
).
11.
N.
Syn
,
L.
Wang
,
G.
Sethi
,
J.-P.
Thiery
, and
B.-C.
Goh
,
Trends Pharmacol. Sci.
37
(
7
),
606
617
(
2016
).
12.
R. C.
Lai
,
F.
Arslan
,
M. M.
Lee
,
N. S.
Sze
,
A.
Choo
,
T. S.
Chen
,
M.
Salto-Tellez
,
L.
Timmers
,
C. N.
Lee
,
R. M.
El Oakley
,
G.
Pasterkamp
,
D. P.
de Kleijn
, and
S. K.
Lim
,
Stem Cell Res.
4
(
3
),
214
222
(
2010
).
13.
J. P.
Sluijter
,
V.
Verhage
,
J. C.
Deddens
,
F.
van den Akker
, and
P. A.
Doevendans
,
Cardiovasc. Res.
102
(
2
),
302
311
(
2014
).
14.
A.
Aharon
,
A.
Rebibo-Sabbah
,
I.
Tzoran
, and
C.
Levin
,
Rambam Maimonides Med. J.
5
(
4
),
e0032
(
2014
).
15.
L.
Muller
,
C. S.
Hong
,
D. B.
Stolz
,
S. C.
Watkins
, and
T. L.
Whiteside
,
J. Immunol. Methods
411
,
55
65
(
2014
).
16.
R.
Kalluri
,
J. Clin. Invest.
126
(
4
),
1208
1215
(
2016
).
17.
F.
Cappello
,
M.
Logozzi
,
C.
Campanella
,
C. C.
Bavisotto
,
A.
Marcilla
,
F.
Properzi
, and
S.
Fais
,
Eur. J. Pharm. Sci.
96
,
93
98
(
2017
).
18.
A.
Panagiotara
,
A.
Markou
,
E. S.
Lianidou
,
G. P.
Patrinos
, and
T.
Katsila
,
Public Health Genomics
20
(
2
),
116
125
(
2017
).
19.
H.
Kalra
,
C. G.
Adda
,
M.
Liem
,
C. S.
Ang
,
A.
Mechler
,
R. J.
Simpson
,
M. D.
Hulett
, and
S.
Mathivanan
,
Proteomics
13
(
22
),
3354
3364
(
2013
).
20.
B. J.
Tauro
,
D. W.
Greening
,
R. A.
Mathias
,
H.
Ji
,
S.
Mathivanan
,
A. M.
Scott
, and
R. J.
Simpson
,
Methods
56
(
2
),
293
304
(
2012
).
21.
R.
Linares
,
S.
Tan
,
C.
Gounou
,
N.
Arraud
, and
A. R.
Brisson
,
J. Extracell. Vesicles
4
(
1
),
29509
(
2015
).
22.
J.
Van Deun
,
P.
Mestdagh
,
R.
Sormunen
,
V.
Cocquyt
,
K.
Vermaelen
,
J.
Vandesompele
,
M.
Bracke
,
O.
De Wever
, and
A.
Hendrix
,
J. Extracell. Vesicles
3
(
1
),
24858
(
2014
).
23.
F.
Yang
,
X.
Liao
,
Y.
Tian
, and
G.
Li
,
Biotechnol. J.
12
(
4
),
1600699
(
2017
).
24.
J. C.
Contreras-Naranjo
,
H.-J.
Wu
, and
V. M.
Ugaz
,
Lab Chip
17
(
21
),
3558
3577
(
2017
).
25.
S. S.
Kanwar
,
C. J.
Dunlay
,
D. M.
Simeone
, and
S.
Nagrath
,
Lab Chip
14
(
11
),
1891
1900
(
2014
).
26.
M.
He
,
J.
Crow
,
M.
Roth
,
Y.
Zeng
, and
A. K.
Godwin
,
Lab Chip
14
(
19
),
3773
3780
(
2014
).
27.
S.
Fang
,
H.
Tian
,
X.
Li
,
D.
Jin
,
X.
Li
,
J.
Kong
,
C.
Yang
,
X.
Yang
,
Y.
Lu
,
Y.
Luo
,
B.
Lin
,
W.
Niu
, and
T.
Liu
,
PLoS One
12
(
4
),
e0175050
(
2017
).
28.
P.
Zhang
,
M.
He
, and
Y.
Zeng
,
Lab Chip
16
(
16
),
3033
3042
(
2016
).
29.
M. S.
Pols
and
J.
Klumperman
,
Exp. Cell Res.
315
(
9
),
1584
1592
(
2009
).
30.
P.
Li
,
M.
Kaslan
,
S. H.
Lee
,
J.
Yao
, and
Z.
Gao
,
Theranostics
7
(
3
),
789
804
(
2017
).
31.
I.
Furi
,
F.
Momen-Heravi
, and
G.
Szabo
,
Ann. Transl. Med.
5
(
12
),
263
(
2017
).
32.
H.
Im
,
H.
Shao
,
Y. I.
Park
,
V. M.
Peterson
,
C. M.
Castro
,
R.
Weissleder
, and
H.
Lee
,
Nat. Biotechnol.
32
(
5
),
490
495
(
2014
).
33.
R. T.
Davies
,
J.
Kim
,
S. C.
Jang
,
E.-J.
Choi
,
Y. S.
Gho
, and
J.
Park
,
Lab Chip
12
(
24
),
5202
5210
(
2012
).
34.
K.
Lee
,
H.
Shao
,
R.
Weissleder
, and
H.
Lee
,
ACS Nano
9
(
3
),
2321
2327
(
2015
).
35.
M.
Wu
,
Y.
Ouyang
,
Z.
Wang
,
R.
Zhang
,
P.-H.
Huang
,
C.
Chen
,
H.
Li
,
P.
Li
,
D.
Quinn
,
M.
Dao
,
S.
Suresh
,
Y.
Sadovsky
, and
T. J.
Huang
,
Proc. Natl. Acad. Sci. U.S.A.
114
(
40
),
10584
10589
(
2017
).
36.
C.
Liu
,
J.
Guo
,
F.
Tian
,
N.
Yang
,
F.
Yan
,
Y.
Ding
,
J.
Wei
,
G.
Hu
,
G.
Nie
, and
J.
Sun
,
ACS Nano
11
(
7
),
6968
6976
(
2017
).
37.
S. M.
Santana
,
M. A.
Antonyak
,
R. A.
Cerione
, and
B. J.
Kirby
,
Biomed. Microdevices
16
(
6
),
869
877
(
2014
).
38.
R.
Burger
,
D.
Kirby
,
M.
Glynn
,
C.
Nwankire
,
M.
O'Sullivan
,
J.
Siegrist
,
D.
Kinahan
,
G.
Aguirre
,
G.
Kijanka
,
R. A.
Gorkin
 3rd
, and
J.
Ducree
,
Curr. Opin. Chem. Biol.
16
(
3–4
),
409
414
(
2012
).
39.
R.
Burger
and
J.
Ducrée
,
Expert Rev. Mol. Diagn.
12
(
4
),
407
421
(
2012
).
40.
Z. T. F.
Yu
,
J. G.
Joseph
,
S. X.
Liu
,
M. K.
Cheung
,
P. J.
Haffey
,
K.
Kurabayashi
, and
J.
Fu
,
Sens Actuators B Chem
245
,
1050
1061
(
2017
).
41.
L.
Clime
,
D.
Brassard
,
M.
Geissler
, and
T.
Veres
,
Lab on a Chip
15
(
11
),
2400
2411
(
2015
).
42.
W.
Al-Faqheri
,
F.
Ibrahim
,
T. H.
Thio
,
N.
Bahari
,
H.
Arof
,
H. A.
Rothan
,
R.
Yusof
, and
M.
Madou
,
Sensors
15
(
3
),
4658
4676
(
2015
).
43.
S.
Haeberle
,
T.
Brenner
,
R.
Zengerle
, and
J.
Ducree
,
Lab Chip
6
(
6
),
776
781
(
2006
).
44.
O.
Strohmeier
,
M.
Keller
,
F.
Schwemmer
,
S.
Zehnle
,
D.
Mark
,
F.
von Stetten
,
R.
Zengerle
, and
N.
Paust
,
Chem. Soc. Rev.
44
(
17
),
6187
6229
(
2015
).
45.
J.
Ducrée
,
S.
Haeberle
,
S.
Lutz
,
S.
Pausch
,
F. v.
Stetten
, and
R.
Zengerle
,
J. Micromech. Microeng.
17
(
7
),
S103
S115
(
2007
).
46.
J. H.
Son
,
S. H.
Lee
,
S.
Hong
,
S. M.
Park
,
J.
Lee
,
A. M.
Dickey
, and
L. P.
Lee
,
Lab Chip
14
(
13
),
2287
2292
(
2014
).
47.
J. C.
Yeo
,
Z.
Wang
, and
C. T.
Lim
,
Biomicrofluidics
9
(
5
),
054114
(
2015
).
48.
A.
Lee
,
J.
Park
,
M.
Lim
,
V.
Sunkara
,
S. Y.
Kim
,
G. H.
Kim
,
M. H.
Kim
, and
Y. K.
Cho
,
Anal. Chem.
86
(
22
),
11349
11356
(
2014
).
49.
H.-K.
Woo
,
V.
Sunkara
,
J.
Park
,
T.-H.
Kim
,
J.-R.
Han
,
C.-J.
Kim
,
H.-I.
Choi
,
Y.-K.
Kim
, and
Y.-K.
Cho
,
ACS Nano
11
(
2
),
1360
1370
(
2017
).
50.
J.
Schageman
,
E.
Zeringer
,
M.
Li
,
T.
Barta
,
K.
Lea
,
J.
Gu
,
S.
Magdaleno
,
R.
Setterquist
, and
A. V.
Vlassov
,
BioMed Res. Int.
2013
,
253957
.
51.
S. J.
Gould
and
G.
Raposo
,
J. Extracell. Vesicles
2
(
1
),
20389
(
2013
).
52.
V.
Budnik
,
C.
Ruiz-Cañada
, and
F.
Wendler
,
Nat. Rev. Neurosci.
17
(
3
),
160
172
(
2016
).
53.
V.
Palanisamy
,
S.
Sharma
,
A.
Deshpande
,
H.
Zhou
,
J.
Gimzewski
, and
D. T.
Wong
,
PLoS One
5
(
1
),
e8577
(
2010
).
54.
E. I.
Buzas
,
B.
Gyorgy
,
G.
Nagy
,
A.
Falus
, and
S.
Gay
,
Nat. Rev. Rheumatol.
10
(
6
),
356
364
(
2014
).
55.
T.
Tian
,
Y.
Wang
,
H.
Wang
,
Z.
Zhu
, and
Z.
Xiao
,
J. Cell. Biochem.
111
(
2
),
488
496
(
2010
).
56.
S.
Sharma
and
J. K.
Gimzewski
,
J. Nanomed. Nanotechnol.
3
(
7
),
1000e115
(
2012
).
57.
M. S.
Kwon
,
S.-H.
Shin
,
S.-H.
Yim
,
K. Y.
Lee
,
H.-M.
Kang
,
T.-M.
Kim
, and
Y.-J.
Chung
,
Lung Cancer
57
(
1
),
46
53
(
2007
).

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