Efficiency of applying chicken IgY antibody conjugates against human IgG in ELISA and dot immunoassay

• Alexander G. Poltavchenko
• Anna V. Ersh
• Nikita D. Ushkalenko
• Pavel V. Filatov
• Evgeniia A. Kolosova
• Daniil V. Shanshin
• Dmitry N. Shcherbakov

Abstract

The aim of the study – comparative evaluation of chicken (IgY) and mouse monoclonal (IgG) antibodies against human IgG as a probe for ELISA and dot immunoassay test systems in the diagnosis of tick-borne encephalitis.

Material and methods. Experiments were conducted to compare the ability of conjugates of mouse IgG and chicken IgY antibodies against human IgG, as well as proteins A and G associated with horseradish peroxidase and colloidal gold to determine human immunoglobulins of different classes, as well as to detect IgG in human sera specific to the tick-borne encephalitis virus (TBEV). A recombinant analogue of domain III of TBEV glycoprotein E was used as a capture reagent in ELISA and dot analysis. The studies were performed in comparison with the commercial VectoTBEV-IgG kit using panels of blood sera containing antibodies to tick-borne encephalitis, yellow fever and dengue viruses, as well as samples from healthy donors.

Results and discussion. It was shown that the designed experimental kits for ELISA and dot analysis adequately identify positive and negative samples and allow one to differentiate antibodies to TBEV from antibodies to other flaviviruses. Peroxidase and colloidal gold conjugates provide similar detection limits for IgG, and chicken antibody probes exhibit stronger optical signals and sensitivity advantages over mouse IgG conjugates.

Conclusion. The data obtained allow us to consider conjugates based on chicken antibodies as promising reagents for immunoassays.

Keywords:IgY; IgG; tick-borne encephalitis; conjugates; immunodiagnostics; dot-immunoassay; ELISA

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