Ultrasensitive detection of cancer biomarkers in the clinic by use of a nanostructured microfluidic array.
Ultrasensitive detection of cancer biomarkers in the clinic by use of a nanostructured microfluidic array.
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DOI：
10.1021/ac301392g
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发表时间：
2012-07-17
影响因子：
7.4
通讯作者：
Rusling, James F.
中科院分区：
文献类型：
--
作者：
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.
由于缺乏合适的测量设备和缺乏严格验证的蛋白质组，多重生物标志物蛋白质检测对于临床癌症诊断具有未实现的前景。在这里，我们报告了一种超灵敏的电化学微流体阵列，该阵列经过优化以测量生物标志物蛋白的四种蛋白质组，并且我们验证了该蛋白质组用于准确的口腔癌诊断。在5-50 fg mL−1范围内实现了前所未有的超低检测，可同时测量稀释血清中的蛋白质IL-6、IL-8、VEGF和VEGF-C。该免疫阵列通过使用由携带400，000个酶标记和120，000个抗体的磁珠进行的离线蛋白质捕获，在50分钟测定中实现了高灵敏度。在捕获蛋白质并洗涤以抑制非特异性结合后，珠子被磁性分离并注入阵列中，以通过八个纳米结构传感器上的抗体进行选择性捕获。与ELISA在条件癌细胞培养基中的蛋白质测定的良好相关性证实了这种方法的准确性。在78个口腔癌患者血清样品和49个对照中的4-蛋白水平的标准化平均值对于口腔癌检测给出了89%的临床灵敏度和98%的特异性，证明了高诊断实用性。该免疫芯片成本低、制作简单，为口腔癌的诊断和个体化治疗提供了一种快速的血清学检测方法。该设备很容易适用于其他癌症的临床诊断。
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 IL-6, IL-8, 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 120,000 antibodies. After capturing the proteins and washing to inhibit non-specific binding, the beads are magnetically separated and injected into the array for selective capture by antibodies on eight nanostructured sensors. Good correlations with ELISA for protein determinations in conditioned cancer cell media confirmed the accuracy of this approach. Normalized means of the 4-protein levels in 78 oral cancer patient serum samples and 49 controls gave clinical sensitivity 89% and specificity 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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影响因子：
12.6
作者：
Chikkaveeraiah, Bhaskara V.;Mani, Vigneshwaran;Patel, Vyomesh;Gutkind, J. Silvio;Rusling, James F.
通讯作者：
Rusling, James F.
影响因子：
17.1
作者：
Mani, Vigneshwaran;Chikkaveeraiah, Bhaskara V.;Patel, Vyomesh;Gutkind, J. Silvio;Rusling, James F.
通讯作者：
Rusling, James F.
影响因子：
4.8
作者：
Benke, Emily M.;Ji, Youngmi;Yeudall, W. Andrew
通讯作者：
Yeudall, W. Andrew
影响因子：
3.7
作者：
Castilho RM;Squarize CH;Leelahavanichkul K;Zheng Y;Bugge T;Gutkind JS
通讯作者：
Gutkind JS
影响因子：
16.6
作者：
Krishnan, Sadagopan;Mani, Vigneshwaran;Wasalathanthri, Dhanuka;Kumar, Challa V.;Rusling, James F.
通讯作者：
Rusling, James F.