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A noninstrumental immunoassay based on colloidal dyes

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.

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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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Correspondence to I. A. Lubavina.

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Lubavina, I.A., Salomatina, I.S., Zinchenko, A.A. et al. A noninstrumental immunoassay based on colloidal dyes. Russ J Bioorg Chem 26, 207–212 (2000). https://doi.org/10.1007/BF02786347

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  • DOI: https://doi.org/10.1007/BF02786347

Key words

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