Feasibility of asymmetrical flow field-flow fractionation as a method for detecting protective antigen by direct recognition of size-increased target-captured nanoprobes

Kayeong Shin, Jaeyeong Choi, Jun Haeng Cho, Moon-Young Yoon, Seungho Lee, Hoeil Chung

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Asymmetrical flow field-flow fractionation (AF4) was evaluated as a potential analytical method for detection of a protective antigen (PA), an Anthrax biomarker. The scheme was based on the recognition of altered AF4 retention through the generation of the size-increased Au nanoparticle probes as a result of PA binding, in which a PA-selective peptide was conjugated on the probe surface. In the visible absorption-based AF4 fractograms, the band position shifted to a longer retention time as the PA concentration increased due to the presence of probe bound with PAs. The shift was insignificant when the concentration was relatively low at 84.3. pM. To improve sensitivity, two separate probes conjugated with two different peptides able to bind on different PA epitopes were used together. The band shift then became distinguishable even at 84.3. pM of PA sample. The formation of larger PA-probe inter-connected species using the dual-probe system was responsible for the enhanced band shift. In parallel, the feasibility of surface-enhanced Raman scattering (SERS) as a potential AF4 detection method was also evaluated. In the off-line SERS fractogram constructed using fractions collected during AF4 separation, a band shift was also observed for the 84.3. pM PA sample, and the band intensity was higher when using the dual-probe system. The combination of AF4 and SERS is promising for the detection of PA and will become a potential tool if the reproducibility of SERS measurement is improved.

Original languageEnglish
Pages (from-to)239-246
Number of pages8
JournalJournal of Chromatography A
Volume1422
DOIs
StatePublished - 2015 Nov 27

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Field Flow Fractionation
Nanoprobes
Fractionation
Flow fields
Antigens
Raman Spectrum Analysis
Raman scattering
Peptides
Biomarkers
Nanoparticles
Epitopes

Keywords

  • Anthrax
  • Asymmetrical flow field-flow fractionation (AF4)
  • Protective antigen (PA)
  • Surface-enhanced Raman scattering (SERS)

Cite this

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title = "Feasibility of asymmetrical flow field-flow fractionation as a method for detecting protective antigen by direct recognition of size-increased target-captured nanoprobes",
abstract = "Asymmetrical flow field-flow fractionation (AF4) was evaluated as a potential analytical method for detection of a protective antigen (PA), an Anthrax biomarker. The scheme was based on the recognition of altered AF4 retention through the generation of the size-increased Au nanoparticle probes as a result of PA binding, in which a PA-selective peptide was conjugated on the probe surface. In the visible absorption-based AF4 fractograms, the band position shifted to a longer retention time as the PA concentration increased due to the presence of probe bound with PAs. The shift was insignificant when the concentration was relatively low at 84.3. pM. To improve sensitivity, two separate probes conjugated with two different peptides able to bind on different PA epitopes were used together. The band shift then became distinguishable even at 84.3. pM of PA sample. The formation of larger PA-probe inter-connected species using the dual-probe system was responsible for the enhanced band shift. In parallel, the feasibility of surface-enhanced Raman scattering (SERS) as a potential AF4 detection method was also evaluated. In the off-line SERS fractogram constructed using fractions collected during AF4 separation, a band shift was also observed for the 84.3. pM PA sample, and the band intensity was higher when using the dual-probe system. The combination of AF4 and SERS is promising for the detection of PA and will become a potential tool if the reproducibility of SERS measurement is improved.",
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Feasibility of asymmetrical flow field-flow fractionation as a method for detecting protective antigen by direct recognition of size-increased target-captured nanoprobes. / Shin, Kayeong; Choi, Jaeyeong; Cho, Jun Haeng; Yoon, Moon-Young; Lee, Seungho; Chung, Hoeil.

In: Journal of Chromatography A, Vol. 1422, 27.11.2015, p. 239-246.

Research output: Contribution to journalArticle

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T1 - Feasibility of asymmetrical flow field-flow fractionation as a method for detecting protective antigen by direct recognition of size-increased target-captured nanoprobes

AU - Shin, Kayeong

AU - Choi, Jaeyeong

AU - Cho, Jun Haeng

AU - Yoon, Moon-Young

AU - Lee, Seungho

AU - Chung, Hoeil

PY - 2015/11/27

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