A rapid method to regenerate piezoelectric microcantilever sensors (PEMS).

A rapid method to regenerate piezoelectric microcantilever sensors (PEMS).
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DOI:
10.3390/s110505520
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发表时间:
2011
期刊:
Sensors (Basel, Switzerland)
影响因子:
--
通讯作者:
Adams GP
Adams GP
中科院分区:
其他
文献类型:
--
作者:
Loo L;Wu W;Shih WY;Shih WH;Borghaei H;Pourrezaei K;Adams GP

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压电微悬臂梁传感器(PEMS)是一种用于检测生物体液中蛋白质和细胞的灵敏工具。然而,目前可用的PEMS只能使用一次,或者在后续使用之前必须完全剥离和再功能化。在这里,我们报告了一个替代的再生协议,采用高盐浓度,以消除目标,留下固定在微悬臂梁表面的功能探针的成功使用。我们的模型系统采用重组人表皮生长因子受体(EGFR)的胞外结构域(ECD)作为探针,抗人EGFR多克隆抗体作为靶标。我们报告,高浓度的氯化镁解离的多克隆抗体特异性结合到EGFR ECD固定在传感器表面上,而不影响其生物活性。这种简单的再生方案既最小化了重新缀合探针所需的时间,又保持了固定在PEMS表面上的探针的密度,从而在一系列测定中产生相同的生物传感器灵敏度。
Piezoelectric microcantilever sensors (PEMS) can be sensitive tools for the detection of proteins and cells in biological fluids. However, currently available PEMS can only be used a single time or must be completely stripped and refunctionalized prior to subsequent uses. Here we report the successful use of an alternative regeneration protocol employing high salt concentrations to remove the target, leaving the functional probe immobilized on the microcantilever surface. Our model system employed the extracellular domain (ECD) of recombinant human Epidermal Growth Factor Receptor (EGFR) as the probe and anti-human EGFR polyclonal antibodies as the target. We report that high concentrations of MgCl2 dissociated polyclonal antibodies specifically bound to EGFR ECD immobilized on the sensor surface without affecting its bioactivity. This simple regeneration protocol both minimized the time required to re-conjugate the probe and preserved the density of probe immobilized on PEMS surface, yielding identical biosensor sensitivity over a series of assays.
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