The selection and optimization of the detection system for self-contained multiplexed dot-immunoassay

Aleksandr Georgievich Poltavchenko, Boris Nikolaevich Zaitsev, Anna Vasilievna Ersh, Denis Vladimirovich Korneev, Oleg Sviatoslavovich Taranov, Pavel Vladimirovich Filatov, Oleg Viacheslavovich Nechitaylo

Research output: Contribution to journalArticleResearchpeer-review

7 Citations (Scopus)

Abstract

This study performed a comparative estimation of the detection systems with the use of conjugates based on peroxidase, alkaline phosphatase, colloidal gold, and the amplification system “Super-CARD” for multiplex dot-immunoassay of antibodies. The results of the study show that the sensitivity of the detection system with colloidal gold was approximately 8 times higher than that of the system with amplification “Super-CARD”, 30 times higher than that of the system with conjugate of alkaline phosphatase, and 250 times higher than the sensitivity of the system with the peroxidase conjugate. Gold immunosols limit the direct detection of human IgG of 10 pg with dynamic range of optical signal change from 5 ng to 10 pg. This limit corresponds to a range of concentrations of IgG from 2.5 μg/mL to 5 ng/mL and covers the range of specific IgG concentrations, to which a typical natural immune response leads. The most typical reasons for aggregate formation during obtainment of colloidal gold and the binding of colloidal gold with biocomponents were explored. The method of minimization of particle clusters formation was suggested as well as the method for increasing stability of diluted preparations of probe by means of sedimentation of aggregates from the ready-made product.

Original languageEnglish
Pages (from-to)540-554
Number of pages15
JournalJournal of Immunoassay and Immunochemistry
Volume37
Issue number5
DOIs
Publication statusPublished - 2 Sep 2016
Externally publishedYes

Keywords

  • aggregation
  • colloidal gold
  • immunochemical probe
  • multiplexed dot-immunoassay
  • sensitivity
  • stability

Cite this