Genetically functionalized ferritin nanoparticles with a high-affinity protein binder for immunoassay and imaging

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Molecular detection of target molecules with high sensitivity and specificity is of great significance in bio and medical sciences. Here, we present genetically functionalized ferritin nanoparticles with a high-affinity protein binder, and their utility as a signal generator in a variety of immunoassays and imaging. As a high-affinity protein binder, human IgG-specific repebody, which is composed of LRR (Leucinerich repeat) modules, was used. The repebody was genetically fused to the N-terminal heavy-chain ferritin, and the resulting subunits were self-assembled to the repebody-ferritin nanoparticles composed of 24 subunits. The repebody-ferritin nanoparticles were shown to have a three-order of magnitude higher binding affinity toward human IgG than free repebody mainly owing to a decreased dissociation rate constant. The repebody-ferritin nanoparticles were conjugated with fluorescent dyes, and the resulting nanoparticles were used for western blotting, cell imaging, and flow cytometric analysis. The dye-labeled repebody-ferritin nanoparticles were shown to generate about 3-fold stronger fluorescent signals in immunoassays than monovalent repebody. The repebody-functionalized ferritin nanoparticles can be effectively used for sensitive and specific immunoassays and imaging in many areas. (C) 2017 Elsevier B.V. All rights reserved.
Publisher
ELSEVIER SCIENCE BV
Issue Date
2017-10
Language
English
Article Type
Article
Keywords

CARBOHYDRATE ANTIGEN 125; SIGNAL AMPLIFICATION; DRUG-DELIVERY; ELECTROCHEMICAL IMMUNOASSAY; TUMOR-CELLS; H-CHAIN; BINDING; APOFERRITIN; CANCER; EXPRESSION

Citation

ANALYTICA CHIMICA ACTA, v.988, pp.81 - 88

ISSN
0003-2670
DOI
10.1016/j.aca.2017.07.060
URI
http://hdl.handle.net/10203/226277
Appears in Collection
BS-Journal Papers(저널논문)
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