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Multiparameter single-particle motion analysis for homogeneous digital immunoassay.

Kenji Akama1, Hiroyuki Noji

  • 1Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan. hnoji@g.ecc.u-tokyo.ac.jp.

The Analyst
|December 28, 2020
PubMed
Summary

This study introduces a refined digital homogeneous non-enzymatic immunoassay (digital Ho-Non ELISA) for highly sensitive biomolecule quantification. By analyzing nanoparticle motion parameters, researchers significantly improved assay specificity and sensitivity, enhancing the potential of digital immunoassays.

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Area of Science:

  • Biotechnology
  • Analytical Chemistry
  • Nanotechnology

Background:

  • Digital homogeneous non-enzymatic immunoassays (digital Ho-Non ELISA) offer sensitive biomolecule quantification.
  • Current methods face challenges in discriminating specifically bound nanoparticles from non-specifically immobilized ones, impacting specificity.

Purpose of the Study:

  • To enhance the specificity of digital Ho-Non ELISA by analyzing multiple nanoparticle motion parameters.
  • To improve the discrimination between target-bound and non-specifically immobilized nanoparticles.

Main Methods:

  • Developed a multiparameter single-particle motion analysis approach.
  • Corrected root-mean-square displacement (RMSD) using bead fluorescence intensity to account for bead size heterogeneity.
  • Classified nanoparticle motion patterns using the aspect ratio of their x-y trajectories.

Main Results:

  • Achieved a 3.9-fold enhancement in sensitivity for Prostate-Specific Antigen (PSA) assay compared to conventional RMSD analysis.
  • Successfully discriminated between specifically tethered and non-specifically immobilized particles by analyzing corrected RMSD and aspect ratio.
  • Demonstrated improved specificity in the digital immunoassay.

Conclusions:

  • Multiparameter analysis of nanoparticle motion significantly enhances the specificity and sensitivity of digital immunoassays.
  • This refined digital Ho-Non ELISA strategy broadens the potential applications of digital immunoassay technology.
  • The method provides a robust approach for accurate biomolecule quantification.