Evaluation of click chemistry microarrays for immunosensing of alpha-fetoprotein (AFP)
Evaluation of click chemistry microarrays for immunosensing of alpha-fetoprotein (AFP)
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DOI：
10.3762/bjnano.10.241
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发表时间：
2019-12
影响因子：
3.1
通讯作者：
Seyed Mohammad Mahdi Dadfar;S. Sekula-Neuner;V. Trouillet;Hui-yu Liu;Ravi Kumar;A. Powell;Michael Hirtz
中科院分区：
文献类型：
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作者：
Seyed Mohammad Mahdi Dadfar;S. Sekula-Neuner;V. Trouillet;Hui-yu Liu;Ravi Kumar;A. Powell;Michael Hirtz
细胞、组织或体液中癌症生物标志物的水平可用于预测癌症的存在，甚至可以指示疾病的阶段。甲胎蛋白（AFP）是肝细胞癌（HCC）早期筛查和诊断最常用的生物标志物。在这里，结合了三种技术（点击化学、生物素-链霉亲和素-生物素夹心策略以及抗原-抗体相互作用的使用）来实现用于 AFP 检测的灵敏荧光免疫传感器。三种类型的功能化玻璃（二苯并环辛炔-（DBCO-）、硫醇和环氧基封端表面）通过使用各自适当的点击化学对应物（第一类是生物素-硫醇或生物素-叠氮化物，第二类是生物素-马来酰亚胺或生物素-DBCO，以及生物素-胺或生物素-硫醇）进行生物素化。 对于第三类）。通过生物素-链霉亲和素-生物素夹心技术将抗 AFP 抗体固定在表面。为了评估不同制备表面的传感性能，通过微通道悬臂点样 (μCS) 将荧光标记的 AFP 点样到这些表面上。根据荧光测量，找到了最佳的微阵列设计并确定了其灵敏度。
The level of cancer biomarkers in cells, tissues or body fluids can be used for the prediction of the presence of cancer or can even indicate the stage of the disease. Alpha-fetoprotein (AFP) is the most commonly used biomarker for early screening and diagnosis of hepatocellular carcinoma (HCC). Here, a combination of three techniques (click chemistry, the biotin–streptavidin–biotin sandwich strategy and the use of antigen–antibody interactions) were combined to implement a sensitive fluorescent immunosensor for AFP detection. Three types of functionalized glasses (dibenzocyclooctyne- (DBCO-), thiol- and epoxy-terminated surfaces) were biotinylated by employing the respective adequate click chemistry counterparts (biotin–thiol or biotin–azide for the first class, biotin–maleimide or biotin–DBCO for the second class and biotin–amine or biotin–thiol for the third class). The anti-AFP antibody was immobilized on the surfaces via a biotin–streptavidin–biotin sandwich technique. To evaluate the sensing performance of the differently prepared surfaces, fluorescently labeled AFP was spotted onto them via microchannel cantilever spotting (µCS). Based on the fluorescence measurements, the optimal microarray design was found and its sensitivity was determined.