Ultrasensitive detection of cancer biomarkers in the clinic by use of a nanostructured microfluidic array

Malhotra, R. and Patel, V. and Chikkaveeraiah, B.V. and Munge, B.S. and Cheong, S.C. and Zain, R.B. and Abraham, M.T. and Dey, D.K. and Gutkind, J.S. and Rusling, J.F. (2012) Ultrasensitive detection of cancer biomarkers in the clinic by use of a nanostructured microfluidic array. Analytical Chemistry, 84 (14). pp. 6249-6255. ISSN 0003-2700, DOI https://doi.org/10.1021/ac301392g.


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Official URL: http://www.ncbi.nlm.nih.gov/pubmed/22697359


Multiplexed biomarker protein detection holds unrealized promise for clinical cancer diagnostics due to lack of suitable measurement devices and lack of rigorously validated protein panels. Here we report an ultrasensitive electrochemical microfluidic array optimized to measure a four-protein panel of biomarker proteins, and we validate the protein panel for accurate oral cancer diagnostics. Unprecedented ultralow detection into the 5-50 fg.mL(-1) range was achieved for simultaneous measurement of proteins interleukin 6 (IL-6), IL-8, vascular endothelial growth factor (VEGF), and VEGF-C in diluted serum. The immunoarray achieves high sensitivity in 50 min assays by using off-line protein capture by magnetic beads carrying 400 000 enzyme labels and similar to 100 000 antibodies. After capture of the proteins and washing to inhibit nonspecific binding, the beads are magnetically separated and injected into the array for selective capture by antibodies on eight nanostructured sensors. Good correlations with enzyme-linked immunosorbent assays (ELISA) for protein determinations in conditioned cancer cell media confirmed the accuracy of this approach. Normalized means of the four protein levels in 78 oral cancer patient serum samples and 49 controls gave clinical sensitivity of 89 and specificity of 98 for oral cancer detection, demonstrating high diagnostic utility. The low-cost, easily fabricated immunoarray provides a rapid Serum test for diagnosis and personalized therapy of oral cancer. The device is readily adaptable to clinical diagnostics of other cancers.

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Additional Information: ISI Document Delivery No.: 974NB Times Cited: 1 Cited Reference Count: 46 Cited References: Adler M, 2008, ANALYST, V133, P702, DOI 10.1039/b718587c Benke EM, 2010, ORAL ONCOL, V46, pE19, DOI 10.1016/j.oraloncology.2010.02.006 Castilho RM, 2010, PLOS ONE, V5, DOI 10.1371/journal.pone.0010503 Chikkaveeraiah BV, 2009, ANAL CHEM, V81, P9129, DOI 10.1021/ac9018022 Chikkaveeraiah BV, 2011, BIOSENS BIOELECTRON, V26, P4477, DOI 10.1016/j.bios.2011.05.005 Choong N, 2008, CA-CANCER J CLIN, V58, P32, DOI 10.3322/CA.2007.0004 Wang TB, 2007, WORLD J GASTROENTERO, V13, P1794 Fan R, 2008, NAT BIOTECHNOL, V26, P1373, DOI 10.1038/nbt.1507 GALLATI H, 1979, J CLIN CHEM CLIN BIO, V17, P1 Gaster RS, 2009, NAT MED, V15, P1327, DOI 10.1038/nm.2032 Giljohann DA, 2009, NATURE, V462, P461, DOI 10.1038/nature08605 Gokhale AS, 2005, ORAL ONCOL, V41, P70, DOI 10.1016/j.oraloncology.2004.06.005 Goodsaid FM, 2010, SCI TRANSL MED, V2, DOI 10.1126/scitranslmed.3001040 Hong DS, 2007, CANCER, V110, P1911, DOI 10.1002/cncr.22999 Horowitz AM, 2001, J AM DENT ASSOC, V132, p36S Hu S, 2008, CLIN CANCER RES, V14, P6246, DOI 10.1158/1078-0432.CCR-07-5037 Jang A., 1997, CLIN EXP METASTAS, V15, P468 Jeon GA, 2004, INT J CANCER, V112, P249, DOI 10.1002/ijc.20399 Kingsmore SF, 2006, NAT REV DRUG DISCOV, V5, P310, DOI 10.1038/nrd2006 Kory T., 2009, ANAL CHEM, V81, P5777 Krishnan S, 2011, ANGEW CHEM INT EDIT, V50, P1175, DOI 10.1002/anie.201005607 Lee HJ, 2008, ANALYST, V133, P975, DOI 10.1039/b717527b Li Y, 2004, CLIN CANCER RES, V10, P8442, DOI 10.1158/1078-0432.CCR-04-1167 Ludwig JA, 2005, NAT REV CANCER, V5, P845, DOI 10.1038/nrc1739 Malhotra R, 2010, ANAL CHEM, V82, P3118, DOI 10.1021/ac902802b Malhotra R, 2010, LANGMUIR, V26, P15050, DOI 10.1021/la102306z Mani V, 2009, ACS NANO, V3, P585, DOI 10.1021/nn800863w Mitsuhashi A, 2005, CANCER, V103, P724, DOI 10.1002/cncr.20819 Munge BS, 2011, ANGEW CHEM INT EDIT, V50, P7915, DOI 10.1002/anie.201102941 Nam JM, 2003, SCIENCE, V301, P1884, DOI 10.1126/science.1088755 Riedel F, 2005, ANTICANCER RES, V25, P2761 Rissin DM, 2010, NAT BIOTECHNOL, V28, P595, DOI 10.1038/nbt.1641 Rusling J.F., 2011, ANALYST, V135, P2496 Schneider A, 2008, NEOPLASIA, V10, P1295, DOI 10.1593/neo.08586 Shang ZJ, 2002, INT J ORAL MAX SURG, V31, P495, DOI 10.1054/ijom.2002.0284 Siegel R, 2011, CA-CANCER J CLIN, V61, P212, DOI 10.3322/caac.20121 Siriwardena BSMS, 2008, J CLIN PATHOL, V61, P103, DOI 10.1136/jcp.2007.047662 Sriuranpong V, 2003, CANCER RES, V63, P2948 Thompson Lester, 2006, Ear Nose Throat J, V85, P74 WARSINKE A, 2000, J ANAL CHEM+, V366, P622 Wei F, 2009, CLIN CANCER RES, V15, P4446, DOI 10.1158/1078-0432.CCR-09-0050 Weigum SE, 2010, CANCER PREV RES, V3, P518, DOI 10.1158/1940-6207.CAPR-09-0139 Xu L, 1999, CANCER RES, V59, P5822 YAN SF, 1995, J BIOL CHEM, V270, P11463, DOI 10.1074/jbc.270.19.11463 Yu X, 2006, J AM CHEM SOC, V128, P11199, DOI 10.1021/ja062117e ZWEIG MH, 1993, CLIN CHEM, V39, P561 Malhotra, Ruchika Patel, Vyomesh Chikkaveeraiah, Bhaskara V. Munge, Bernard S. Cheong, Sok Ching Zain, Rosnah B. Abraham, Mannil T. Dey, Dipak K. Gutkind, J. Silvio Rusling, James F. NIH from National Institute of Environmental Health Sciences R01ES013557; NIH from National Institute of Biomedical Imaging and Bioengineering R01EB014586; National Institute of Dental and Craniofacial Research; Ministry of Higher Education, Malaysia UM.C/HIR/MOHE/DENT/06 This work was supported by NIH Grants R01ES013557 from National Institute of Environmental Health Sciences and R01EB014586 from National Institute of Biomedical Imaging and Bioengineering and by the Intramural Research Program, National Institute of Dental and Craniofacial Research. We thank the staff at the Oral Cancer Research & Coordinating Centre, University of Malaya (OCRCC-UM), for coordinating sample acquisition from the MOCDTBS, funded by the Ministry of Higher Education, Malaysia (Project UM.C/HIR/MOHE/DENT/06). Amer chemical soc Washington
Uncontrolled Keywords: Squamous-cell carcinoma endothelial growth-factor oral-cancer vegf-c factor receptor head neck serum proteins interleukin-6
Subjects: R Medicine > RK Dentistry
Divisions: Faculty of Dentistry > Dept of Oral Pathology & Oral Medicine & Periodontology
Depositing User: Mr Ahmad Azwan Azman
Date Deposited: 07 May 2013 03:21
Last Modified: 07 May 2013 03:21
URI: http://eprints.um.edu.my/id/eprint/6027

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