Fluorescent ferritin nanoparticles and application to the aptamer sensor

Seong Eun Kim, Keum Young Ahn, Jin Seung Park, Kyung Rim Kim, Kyung Eun Lee, Sung Sik Han, Jeewon Lee

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

We synthesized fluorescent ferritin nanoparticles (FFNPs) through bacterial expression of the hybrid gene consisting of human ferritin heavy chain (hFTN-H), spacer (glycine-rich peptide), and enhanced green (or red) fluorescent protein [eGFP (or DsRed)] genes. The self-assembly activity of hFTN-H that leads to the formation of nanoparticles (12 nm in diameter), the conformational flexibility of the C-terminus of hFTN-H, and the glycine-rich spacer enabled eGFPs (or DsReds) to be well displayed on the surface of each ferritin nanoparticle, resulting in the construction of green (or red) FFNPs [gFFNPs (or rFFNPs)]. As compared to eGFP (or DsRed) alone, it is notable that the developed FFNPs showed significantly amplified fluorescence intensity and also enhanced stability. DNA aptamers were chemically conjugated to gFFNP via each eGFP's cysteine residue that was newly introduced through site-directed mutagenesis (Ser175Cys). The DNA-aptamer-conjugated gFFNPs were used as a fluorescent reporter probe in the aptamer-based "sandwich" assay of a cancer marker [i.e., platelet-derived growth factor B-chain homodimer (PDGF-BB)] in phosphate-buffered saline buffer or diluted human serum. This is a simple two-step assay without any additional steps for signal amplification, showing that compared to the same aptamer-based assays using eGFP alone or Cy3, the detection signals, affinity of the reporter probe to the cancer marker, and assay sensitivity were significantly enhanced; i.e., the limit of detection was lowered to the 100 fM level. Although the PDGF-BB assay is reported here as a proof-of-concept, the developed FFNPs can be applied in general to any aptamer-based sandwich assays.

Original languageEnglish
Pages (from-to)5834-5843
Number of pages10
JournalAnalytical Chemistry
Volume83
Issue number15
DOIs
Publication statusPublished - 2011 Aug 1

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Ferritins
Assays
Apoferritins
Nanoparticles
Sensors
Nucleotide Aptamers
Glycine
Genes
Proto-Oncogene Proteins c-sis
Mutagenesis
Signal detection
Self assembly
Cysteine
Amplification
Buffers
Fluorescence
Phosphates
Peptides
fluorescent protein 583

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Fluorescent ferritin nanoparticles and application to the aptamer sensor. / Kim, Seong Eun; Ahn, Keum Young; Park, Jin Seung; Kim, Kyung Rim; Lee, Kyung Eun; Han, Sung Sik; Lee, Jeewon.

In: Analytical Chemistry, Vol. 83, No. 15, 01.08.2011, p. 5834-5843.

Research output: Contribution to journalArticle

Kim, Seong Eun ; Ahn, Keum Young ; Park, Jin Seung ; Kim, Kyung Rim ; Lee, Kyung Eun ; Han, Sung Sik ; Lee, Jeewon. / Fluorescent ferritin nanoparticles and application to the aptamer sensor. In: Analytical Chemistry. 2011 ; Vol. 83, No. 15. pp. 5834-5843.
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