Functionalized gold nanoparticles with zinc finger-fused proteins as a colorimetric immunoassay platform

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The quest for highly sensitive and specific detection of disease biomarkers is high, despite many advances in analysis system. Here, we present a sensitive immunoassay platform using DNA-tethered gold nanoparticles and DNA-binding zinc fingers (ZFs). Monomeric alkaline phosphatase (mAP) and human TNF-alpha were employed as a signal generator and a disease biomarker, respectively. Gold nanoparticles (AuNPs) were first grafted with double-stranded DNAs having specific sequences for two different types of ZFs (QNK and zif268). The alkaline phosphatase and TNF-alpha-specific protein binder were genetically fused to each of two different types of ZFs, respectively, followed by conjugation with the DNA-tethered AuNPs in a sequence-specific manner. The use of the functionalized AuNPs as a signal generator in a colorimetric immunoassay of TNF-alpha led to LOD of 120 pg/ml, showing about 161-fold higher sensitivity than a protein binder-fused mAP. The present immunoassay platform could be applied to other analytes by simply replacing a targeting moiety, allowing a versatile and reproducible colorimetric immunoassay.
Publisher
ELSEVIER
Issue Date
2020-08
Language
English
Article Type
Article
Citation

ANALYTICA CHIMICA ACTA, v.1126, pp.154 - 162

ISSN
0003-2670
DOI
10.1016/j.aca.2020.06.019
URI
http://hdl.handle.net/10203/276075
Appears in Collection
BS-Journal Papers(저널논문)
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