Journal
BEILSTEIN JOURNAL OF NANOTECHNOLOGY
Volume 10, Issue -, Pages 2505-2515Publisher
BEILSTEIN-INSTITUT
DOI: 10.3762/bjnano.10.241
Keywords
alpha-fetoprotein (AFP); cancer biomarker; click chemistry; fluorescent immunosensor; hepatocellular carcinoma
Funding
- Karlsruhe Nano Micro Facility (KNMF), a Helmholtz Research Infrastructure at Karlsruhe Institute of Technology (KIT)
- Federal Ministry of Economics and Technology
- Deutsche Forschungsgemeinschaft (DFG) [HI 1724/3-1]
- German Academic Exchange Service (DAAD)
- Karlsruhe House of Young Scientists (KHYS)
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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 (mu CS). Based on the fluorescence measurements, the optimal microarray design was found and its sensitivity was determined.
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