Elaboration of Immunoenzymatic Test-Kit for Total Human IgE Assay and Investigation of Its Analytical Properties
International Journal of Immunology
Volume 1, Issue 1, April 2013, Pages: 1-6
Received: Mar. 3, 2013; Published: Apr. 2, 2013
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Authors
Galkin A. Yu., Department of Industrial Biotechnology, National Technical University of Ukraine “Kyiv Polytechnic Institute”, Kyiv, Ukraine
Dugan A. M., Department of Industrial Biotechnology, National Technical University of Ukraine “Kyiv Polytechnic Institute”, Kyiv, Ukraine
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Abstract
Enzyme-linked immunosorbent assay (ELISA) test-kit has been developed based on complex immuno-chemical, including epitop mapping, characteristics of monoclonal antibodies (MAbs) to human IgЕ. “Sandwich” type ELISA based on usage of different epitop directionality MAbs. It has been founded correlation between affinity of different MAb pairs and its sorbtion-detection ability. Optimal configuration of MAbs in “sandwich” ELISA was follow-ing: 164H10 - 165C12 and 164H10 - 166B7. Cooperative usage of horseradish peroxidase conjugates of MAbs directed to different epitops (165C12-HRP + 166B7-HRP) increased analytical sensitivity of assay and constituted 4.5 IU/ml. Analytical characteristics of developed test-kit were following: dynamic range – from 4.5 to 1600 IU/ml, variation coefficient value during one procedure – 4.2±2.1%, and between procedures – 4.8±2.3%. Presences of human IgG, IgA, IgМ, albumin (1 μg/ml) were not effect on the assay specificity (test concentration of IgE – 50 IU/ml).
Keywords
Human IgE, Monoclonal Antibodies, Immunoenzymatic Analysis
To cite this article
Galkin A. Yu., Dugan A. M., Elaboration of Immunoenzymatic Test-Kit for Total Human IgE Assay and Investigation of Its Analytical Properties, International Journal of Immunology. Vol. 1, No. 1, 2013, pp. 1-6. doi: 10.11648/j.iji.20130101.11
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