Static micro-array isolation, dynamic time series classification, capture and enumeration of spiked breast cancer cells in blood: the nanotube-CTC chip.
We demonstrate the rapid and label-free capture of breast cancer cells spiked in blood using nanotube-antibody micro-arrays. 76-element single wall carbon nanotube arrays were manufactured using photo-lithography, metal deposition, and etching techniques. Anti-epithelial cell adhesion molecule (anti-EpCAM), Anti-human epithelial growth factor receptor 2 (anti-Her2) and non-specific IgG antibodies were functionalized to the surface of the nanotube devices using 1-pyrene-butanoic acid succinimidyl ester. Following device functionalization, blood spiked with SKBR3, MCF7 and MCF10A cells (100/1000 cells per 5 μl per device, 170 elements totaling 0.85 ml of whole blood) were adsorbed on to the nanotube device arrays. Electrical signatures were recorded from each device to screen the samples for differences in interaction (specific or non-specific) between samples and devices. A zone classification scheme enabled the classification of all 170 elements in a single map. A kernel-based statistical classifier for the 'liquid biopsy' was developed to create a predictive model based on dynamic time warping series to classify device electrical signals that corresponded to plain blood (control) or SKBR3 spiked blood (case) on anti-Her2 functionalized devices with ∼90% sensitivity, and 90% specificity in capture of 1000 SKBR3 breast cancer cells in blood using anti-Her2 functionalized devices. Screened devices that gave positive electrical signatures were confirmed using optical/confocal microscopy to hold spiked cancer cells. Confocal microscopic analysis of devices that were classified to hold spiked blood based on their electrical signatures confirmed the presence of cancer cells through staining for DAPI (nuclei), cytokeratin (cancer cells) and CD45 (hematologic cells) with single cell sensitivity. We report 55%-100% cancer cell capture yield depending on the active device area for blood adsorption with mean of 62% (∼12 500 captured off 20 000 spiked cells in 0.1 ml blood) in this first nanotube-CTC chip study.
Small Systems Laboratory, Department of Mechanical Engineering, Worcester Polytechnic Institute, Worcester, MA 01609, USA.
Farhad
F
Khosravi
Patrick J
PJ
Trainor
Christopher
C
Lambert
Goetz
G
Kloecker
Eric
E
Wickstrom
Shesh N
SN
Rai
Balaji
B
Panchapakesan
Clin Chem Lab Med. 2005;43(6):617-27
16006258
Nature. 2007 Dec 20;450(7173):1235-9
18097410
Nanotechnology. 2008 Nov 19;19(46):465101
21836232
Blood. 1994 Apr 1;83(7):1892-902
8142656
J Clin Oncol. 1994 Apr;12(4):725-9
7512130
Front Physiol. 2013 Jul 17;4:185
23882223
Proc Natl Acad Sci U S A. 2009 Jul 28;106(30):12459-64
19620715
J Am Chem Soc. 2005 Sep 14;127(36):12484-5
16144384
Dis Markers. 2002;18(4):167-74
12590170
Clin Cancer Res. 2004 May 1;10(9):3020-8
15131038
Proc Natl Acad Sci U S A. 2004 Jul 20;101(29):10501-4
15249663
N Engl J Med. 2001 Mar 15;344(11):783-92
11248153
Nanotechnology. 2014 May 2;25(17):175201
24721979
Proc Natl Acad Sci U S A. 2010 Oct 26;107(43):18392-7
20930119
N Engl J Med. 2004 Aug 19;351(8):781-91
15317891
Clin Chem Lab Med. 2001 Sep;39(9):811-7
11601678
Acta Cytol. 1965 Mar-Apr;9:169-74
14325797
Cancer Res. 1975 Mar;35(3):512-6
1090362
Clin Cancer Res. 1999 Aug;5(8):1950-60
10473071
Clin Cancer Res. 2004 Oct 15;10(20):6897-904
15501967
Nanotechnology. 2016 Apr 1;27(13):13LT02
26901310
Proc SPIE Int Soc Opt Eng. 2012 Oct 10;8460:84600S
27274607
J Am Chem Soc. 2001 Apr 25;123(16):3838-9
11457124
J Gen Physiol. 1943 Sep 20;27(1):37-60
19873371
Adv Mater. 2011 Jan 4;23(1):94-9
20976831
Int J Oncol. 2005 Jul;27(1):49-57
15942643
Breast Cancer Res Treat. 1998;52(1-3):65-77
10066073
Acta Chir Scand Suppl. 1955;201:1-70
14387468
Hum Pathol. 2004 Jan;35(1):122-8
14745734
Open Access Med Stat. 2016;2016(6):21-29
27942497
J Physiol. 1949 Mar 1;108(1):37-77
18128147
Methods. 2013 Dec 1;64(2):144-52
23816790
Cancer. 1995 Jun 1;75(11):2757-61
7743482
Proc Natl Acad Sci U S A. 1998 Apr 14;95(8):4589-94
9539782
Blood. 1995 Mar 15;85(6):1416-34
7888664
J Cell Physiol. 1995 Oct;165(1):177-85
7559799
J Mol Med (Berl). 1999 Oct;77(10):699-712
10606205
Biosens Bioelectron. 2009 Jul 15;24(11):3372-8
19481922
Breast Cancer Res. 2005;7(6):R975-9
16280045