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Comparative study of serum sample preparation methods in aggregation-based plasmonic sensing.

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Summary
This summary is machine-generated.

Protein corona formation hinders nanoparticle applications. Dialysis effectively removes proteins from serum, enabling sensitive detection of aminoglycoside antibiotics using localized surface plasmon resonance assays.

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Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Nanotechnology

Background:

  • Nanoparticle-based colorimetric methods offer high sensitivity and simplicity for clinical applications.
  • Protein corona formation on nanoparticles (NPs) in biological samples limits their utility.
  • Effective protein removal is crucial for reliable NP-based assays in complex matrices.

Purpose of the Study:

  • To compare protein separation methods for eliminating protein corona on NPs.
  • To develop a sensitive and simple therapeutic drug monitoring (TDM) assay for aminoglycoside antibiotics in serum.
  • To evaluate dialysis as a facile and routine protein separation technique.

Main Methods:

  • Comparison of dialysis, ultrafiltration, and phenol:chloroform:isopentanol extraction for protein removal.
  • Localized surface plasmon resonance (LSPR) aggregation assay to assess assay sensitivity.
  • Coomassie blue staining to identify interfering proteins.
  • Measurement of dialysis efficiency and dialysis coefficient for amikacin.

Main Results:

  • Serum proteins significantly reduce the sensitivity of LSPR aggregation assays.
  • Dialysis effectively removed interfering proteins, enabling naked-eye semi-quantification of aminoglycoside antibiotics (amikacin, tobramycin, streptomycin) at the clinical level.
  • Dialysis demonstrated high efficiency and a favorable dialysis coefficient for amikacin, confirming its efficacy.

Conclusions:

  • Dialysis is a fast, efficient, and universally applicable pretreatment method for removing proteins from crude biological samples.
  • This strategy facilitates the development of sensitive plasmonic assays for small molecules in complex biological matrices.
  • The study overcomes a major limitation in applying NP-based assays to clinical diagnostics.