Russian Journal of Bioorganic Chemistry

, Volume 42, Issue 6, pp 655–663 | Cite as

Biospecific nanoparticles for multiplex phosphorescence analysis (PHOSPHAN)

  • D. V. Paramonov
  • T. S. Kostryukova
  • T. A. Bychenkova
  • V. G. Pomelova
  • N. S. Osin
Article

Abstract

We have developed a technology for the production of polymeric nanoparticles containing the incorporated phosphorescent label (europium ions–naphthoyltrifluoroacetone complexes) and streptavidin that is covalently bound on the surface. The aggregation-stable biospecific nanoparticles (40–60 nm in diameter) include up to 2000 molecular tags/particle and retain biological activity and stable phosphorescence for at least 20 months. They can be used in phosphorescence analysis (PHOSPHANTM)-based biochip technology as an effective detector system to record phosphorescence from microzones (microarrays) printed on the well bottoms of standard polystyrene microplates. The creation of a dense monolayer on the surface of a microzone requires up to 108 particles/microarray, or 109 particles/mm2 of area; this is in good agreement with theoretical estimates. The detection limit is as low as 300–400 phosphorescent nanoparticles per a microzone with an area of ~0.1 mm2. It has been demonstrated in the model of thyroid stimulating hormone (TSH) detection in filter paper dried blood that the newly developed detector system is five times more sensitive than the conventional methods of multiplex PHOSPHAN (with Pt-coproporphyrin phosphorescent label) and lanthanide immune fluoroassay (with fluorescent Eu3+ chelate complexes registered in the enhancement solution). The sensitivity of phosphorescent nanoparticle-based detector system is as low as 6.8 × 105 molecules/1.5 μL sample, which corresponds to a TSH concentration of 1.5 × 10–14 M.

Keywords

nanoparticles phosphorescence analysis (PHOSPHAN) microarrays time-resolved luminescence europium complexes Pt coproporphyrin thyroid-stimulating hormone (TSH) 

Abbreviations

BSA

bovine serum albumin

DVB

divinylbenzene

MAA

metacrylic acid

NTA

2-naphthoyl trifluoroacetone

PHOSPHAN

phosphorescence analysis

Pt-CP

Pt-coproporphyrin

SDS

sodium dodecyl sulfate

TOPO

trioctylphosphine oxide

TR-FIA

time-resolved fluoroimmunoassay

TSH

thyroid-stimulating hormone

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Copyright information

© Pleiades Publishing, Ltd. 2016

Authors and Affiliations

  • D. V. Paramonov
    • 1
  • T. S. Kostryukova
    • 1
  • T. A. Bychenkova
    • 2
  • V. G. Pomelova
    • 2
  • N. S. Osin
    • 2
  1. 1.State Research Institute of Biological Instrument EngineeringMoscowRussia
  2. 2.ImmunoscreenMoscowRussia

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