On-line protein capture on magnetic beads for ultrasensitive microfluidic immunoassays of cancer biomarkers.
On-line protein capture on magnetic beads for ultrasensitive microfluidic immunoassays of cancer biomarkers.
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DOI：
10.1016/j.bios.2013.09.054
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发表时间：
2014-03-15
影响因子：
12.6
通讯作者：
Rusling, James F.
中科院分区：
文献类型：
--
作者：
Otieno, Brunah A.;Krause, Colleen E.;Latus, Alina;Chikkaveeraiah, Bhaskara V.;Faria, Ronaldo C.;Rusling, James F.
生物标志物蛋白的准确、灵敏、多重检测为个性化癌症诊断带来了巨大希望。在这里，我们描述了将一种新型在线室结合到模块化微流体免疫阵列中，以捕获由 300,000 个酶标记和 40,000 个抗体衍生的磁珠上的癌症生物标志物蛋白质。捕获室和检测室由加工模具上的 PDMS 制成，不需要光刻。在搅拌捕获室中，通过磁性固定的珠子从血清或其他生物样品中捕获蛋白质分析物。随后将珠子清洗掉样品成分，并将清洗溶液送往废物处理。移除磁铁和阀门切换后，磁珠-蛋白质生物结合物会被送入检测室，并被 8 个抗体修饰的金纳米颗粒薄膜传感器捕获并进行电流检测。免疫测定中的大多数步骤，包括蛋白质捕获、洗涤和测量，都集成到该设备中。在同步检测中，微流控系统对血清中的白细胞介素 6 (IL-6) 和 IL-8 的超低检测限分别为 5 fg mL−1 和 7 fg mL−1。通过测量口腔癌细胞系条件培养基中的 IL-6 和 IL-8 来证明准确性，并与标准 ELISA 表现出良好的相关性。在线捕获室有助于在适用于多重蛋白质检测的半自动化系统中在 30 分钟内实现快速、灵敏、重复的蛋白质分离和测量。
Accurate, sensitive, multiplexed detection of biomarker proteins holds significant promise for personalized cancer diagnostics. Here we describe the incorporation of a novel on-line chamber to capture cancer biomarker proteins on magnetic beads derivatized with 300,000 enzyme labels and 40,000 antibodies into a modular microfluidic immunoarray. Capture and detection chambers are produced from PDMS on machined molds and do not require lithography. Protein analytes are captured from serum or other biological samples in the stirred capture chamber on the beads held in place magnetically. The beads are subsequently washed free of sample components, and wash solutions sent to waste. Removal of the magnet and valve switching sends the magnetic bead-protein bioconjugates into a detection chamber where they are captured on 8 antibody-decorated gold nanoparticle-film sensors and detected amperometrically. Most steps in the immunoassay including protein capture, washing and measurement are incorporated into the device. In simultaneous assays, the microfluidic system gave ultralow detection limits of 5 fg mL−1 for interleukin-6 (IL-6) and 7 fg mL−1 for IL-8 in serum. Accuracy was demonstrated by measuring IL-6 and IL-8 in conditioned media from oral cancer cell lines and showing good correlations with standard ELISAs. The on-line capture chamber facilitates rapid, sensitive, repetitive protein separation and measurement in 30 min in a semi-automated system adaptable to multiplexed protein detection.
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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：
12.6
作者：
Pan Y;Sonn GA;Sin ML;Mach KE;Shih MC;Gau V;Wong PK;Liao JC
通讯作者：
Liao JC
影响因子：
7.4
作者：
Mani, Vigneshwaran;Wasalathanthri, Dhanuka P.;Joshi, Arnit A.;Kumar, Challa V.;Rusling, James F.
通讯作者：
Rusling, James F.
DOI：
10.1039/c0an00204f
发表时间：
2010-10
期刊：
The Analyst
影响因子：
--
作者：
Rusling JF;Kumar CV;Gutkind JS;Patel V
通讯作者：
Patel V