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Russian Journal of Bioorganic Chemistry

, Volume 26, Issue 3, pp 207–212 | Cite as

A noninstrumental immunoassay based on colloidal dyes

  • I. A. Lubavina
  • I. S. Salomatina
  • A. A. Zinchenko
  • A. V. Zherdev
  • B. B. Dzantiev
Article

Abstract

Detecting labels based on water dispersions of colloidal textile dyes were developed that are useful in various analytical and diagnostic test systems for a simple visual assessment of the assay. Colored water-insoluble particles of dyes were used for the sorptional immobilization of streptavidin on their surface. The resulting streptavidin-dye (STR-DYE) complexes possessed a high visualizing capacity and were used for the combined detection of pesticides (simazine and 2,4-dichlorophenoxyacetic acid) by noninstrumental immunoassay (DYE-comb-assay, competitive dot-immunoassay in the comb format). The detection limits and the duration of our DYE-comb-assay (4 ng/ml, 20–25 min), HRP-comb-assay (competitive dot-immunoassay in the comb format using the enzymic conjugate of STR with horseradish peroxidase) (16 ng/ml), and the traditional competitive ELISA (12–16 ng/ml, 1.5 h) were compared. This DYE-comb-assay is simple enough and can be used under field conditions.

Key words

competitive ELISA colored colloidal dyes 2,4-dichlorophenoxyacetic acid dot-immunoassay simazine 

Abbreviations

BB

borate buffer, pH 7.4

BB/OVA

BB containing ovalbumin (2 mg/ml)

Bi-IgG

biotinylated immunoglobulins

2,4-D

2,4-dichlorophenoxyacetic acid

2,4-D-OVA

a polyvalent conjugate of 2,4-D with ovalbumin

HEPES

N-2-hydroxyethylpiperazine-N-(2-ethanesulfonic acid)

HRP

horseradish peroxidase

MOPS

3-(N-morpholino)propanesulfonic acid

OVA

ovalbumin

PBS

phosphate buffered saline, pH 7.4

PBS/OVA

PBS containing ovalbumin (2 mg/ml)

PBST

PBS containing 0.05% Tween-20

PBST/OVA

PBST containing ovalbumin (2 mg/ml)

Sm

simazine

Sm-Ova

a polyvalent conjugate of simazine with ovalbumin

STR

streptavidin

STR-DYE

a complex of colloidal dye with streptavidin

STR-HRP

a conjugate of streptavidin with horseradish peroxidase

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Copyright information

© MAIK “Nauka/Interperiodica” 2000

Authors and Affiliations

  • I. A. Lubavina
    • 1
  • I. S. Salomatina
    • 1
  • A. A. Zinchenko
    • 1
  • A. V. Zherdev
    • 2
  • B. B. Dzantiev
    • 2
  1. 1.Shemyakin-Ovchinnikov Institute of Bioorganic ChemistryRussian Academy of SciencesGSP-7 MoscowRussia
  2. 2.Bach Institute of BiochemistryRussian Academy of SciencesMoscow

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