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Biosensing of Cardiac Biomarkers Using Single Polyaniline Nanowires

机译:使用单条聚苯胺纳米线对心脏生物标志物的生物传感

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摘要

In this thesis, we explore innovative methods of fabricating single polymer nanowires and applying the fabricated single polymer nanowires for biosensors. The fabrication of single polymer nanowire was carried out via electrochemical deposition growth method, which deposits ionized molecules in a pre-patterned nanochannel between two Au electrodes and forms a nanowire. For biosensing application, we employ polyaniline (PANI) in this research since it has advantages of biocompatibility, easy synthesis, and broad range of electrical conductivity. The single PANI nanowires-based biosensor show high sensitivity and good sensing reliability due to the high surface to volume ratio and single nanowires.udUsing the fabricated single PANI nanowires, the biosensors were developed to detect cardiac biomarkers such as myoglobin (Myo), cardiac troponin I (cTnI), creatine kinase-MB (CK-MB), and B-type natriuretic peptide (BNP). The single PANI nanowires are functionalized by surface immobilization method that was developed in our laboratory using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-Hydroxysuccinimde (NHS) to attach monoclonal antibodies (mAbs) of biomarkers without pre-chemical or physical treatments. Lastly, microfluidic devices including channels and solution infusion/withdrawal system were integrated on the biosensor in order to develop an all-in-one biosensor. Our single PANI nanowire-based biosensor is unique in terms of the functionalization on specific area; the mAbs are immobilized on the only surface of PANI nanowires because PANI has superior biocompatibility to mAbs unlike SiO2 layer or Au electrodes. This advantage reduces functionalization steps comparing to inorganic nanowire biosensors and frees the interfered signals from non-nanowire areas. In addition, the microfluidic channel helps achieving stable electrical signals, high sensitivity, safe sample handling, and minimal damage of nanowire.udThe integrated single PANI nanowire biosensors with microfluidic devices detect very low concentrations of Myo (100 pg/mL), cTnI (250 fg/mL), CK-MB (150 fg/mL), and BNP (50 fg/mL) with good specificity. The remarkable specificity value in cardiac biomarker sensing is over 106 fold, that the specificity value is defined as the ratio of [the highest concentration of non-specific protein showing ignorable or non-response signal] to [the lowest concentration of specific protein showing significant signal change], in the test of bovine serum albumin (BSA) or other cardiac markers. The single PANI nanowire biosensors have shown linear sensing profiles along different concentration from hundreds fg/mL to tens ng/mL depending on the mAbs of the specific biomarkers, and exhibit fast response in a few minutes satisfying reference values of Myo, cTnI, CK-MB, and BNP to diagnose heart failure and determine the patient’s stage of disease.
机译:在本文中,我们探索了制造单一聚合物纳米线并将其应用于生物传感器的创新方法。单个聚合物纳米线的制造是通过电化学沉积生长方法进行的,该方法将电离的分子沉积在两个Au电极之间的预图案化纳米通道中,并形成纳米线。对于生物传感应用,我们在本研究中采用聚苯胺(PANI),因为它具有生物相容性,易于合成和广泛的导电性的优点。基于单PANI纳米线的生物传感器由于具有高的表面体积比和单纳米线,因此显示出高灵敏度和良好的传感可靠性。 uded通过使用制造的单PANI纳米线,该生物传感器用于检测心脏生物标记物,例如肌红蛋白(Myo),心脏肌钙蛋白I(cTnI),肌酸激酶-MB(CK-MB)和B型利钠肽(BNP)。单PANI纳米线通过表面固定化方法进行功能化,该方法是在我们的实验室中使用1-乙基-3-(3-二甲基氨基丙基)碳二亚胺(EDC)和N-羟基琥珀酰亚胺(NHS)连接生物标志物的单克隆抗体(mAbs)而无需预先制备的-化学或物理处理。最后,将包括通道和溶液注入/抽取系统在内的微流体装置集成到生物传感器上,以开发多合一的生物传感器。就特定区域的功能化而言,我们的单个基于PANI纳米线的生物传感器是独一无二的。 mAb固定在PANI纳米线的唯一表面上,因为与SiO2层或Au电极不同,PANI具有优于mAb的生物相容性。与无机纳米线生物传感器相比,该优点减少了功能化步骤,并从非纳米线区域释放了受干扰的信号。此外,微流体通道有助于实现稳定的电信号,高灵敏度,安全的样品处理以及对纳米线的最小破坏。 ud带有微流体设备的集成式PANI纳米线生物传感器可检测极低浓度的Myo(100 pg / mL),cTnI( 250 fg / mL),CK-MB(150 fg / mL)和BNP(50 fg / mL)具有良好的特异性。心脏生物标志物感测中的显着特异性值超过106倍,该特异性值定义为[显示可忽略或无应答信号的非特异性蛋白的最高浓度]与[显示显着性信号的最低浓度的特定蛋白的比值]信号变化],用于测试牛血清白蛋白(BSA)或其他心脏标志物。单个PANI纳米线生物传感器已经显示出线性感应曲线,其浓度从数百fg / mL到数十ng / mL不等,具体取决于特定生物标志物的mAb,并且在几分钟内表现出快速响应,可满足Myo,cTnI,CK-的参考值MB和BNP可以诊断心力衰竭并确定患者的疾病阶段。

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    Lee Innam;

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  • 年度 2012
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