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Microchimica Acta

, Volume 176, Issue 1–2, pp 117–122 | Cite as

Homogeneous immunoassay for human IgG using oriented hen egg IgY immobilized on gold sol nanoparticles

  • Hermine E. Yeritsyan
  • Vardan K. GasparyanEmail author
Original Paper

Abstract

Homogeneous immunoassays using (red) gold nanoparticles represent an attractive detection scheme because of the option of photometric readout. We have applied oriented immobilization of hen egg immunoglobulin Y (IgY) on gold nanoparticles when developing a homogeneous immunoassay for human IgG. In oriented immobilization, as opposed to random immobilization, the antigen binding capabilities of the antibodies are retained. It is shown that such immunoassay has significantly better sensitivity in comparison with methods based on conventional immobilization of affinity-purified antibodies. It is also shown that hen egg IgY is better suited than rabbit antibodies, because much more antibody can be immobilized on gold nanoparticles without any destabilization, probably because of the more acidic nature of these antibodies. In addition, hen egg IgY can be supplied in higher quantity and can be prepared more easily than IgG from rabbits. Bleeding and slaughtering of animals is not needed. The assay presented here has a wide detection range (30–500 ng .mL−1) and a limit of detection as low as 30 ng.mL−1 of human IgG.

Figure

Nanoparticles are treated by thiol for formation of monolayer with exposed NH2 groups. IgY molecule is oxidized by periodate for formation of aldehyde group in Fc fragment. Consequent addition of such antibodies to gold nanoparticles results in binding of IgY molecules to gold nanoparticles via Fc fragment providing oriented immobilization.

Keywords

Gold nanoparticle Immunoassay Immunoglobulin Y Oriented immobilization 

Notes

Acknowledgments

Authors acknowledge to Dr. Mariam Mikaelyan for attentive study of manuscript and for language support. We acknowledge to Dr. Hayk Harutyunyan for support in graphical works.

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  1. 1.H. Buniatian Institute of BiochemistryNational Academy of SciencesYerevanRepublic of Armenia

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