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
Views 2948 Downloads 139
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
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).
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.
E. Harlow, D. Lane, Antibodies. A laboratory manual. N.-Y.: Cold Spring Harbor, 1988. pp. 7-23.
K. Ishizaka, T. Ishizaka, M. Hornbrook, "Physico-chemical properties of human reaginic antibody. IV. Presence of a unique immunoglobulin as a carrier of reaginic activity", J. Immunol., vol. 97 (1), pp. 75–85, July 1966.
D. R. Stanworth, J. H. Humphrey, H. Bennich et al., "Specific inhibition of the Prausnitz-Küstner reaction by an atypical human myeloma protein", Lancet, vol. 290 (7511), pp. 330–332, August 1967.
A. J. Bonnin, F. Montealegre, A. Gewurz, "Association of cigarette smoking with elevated serum IgE levels in His-panic Puerto Rican men", Ann. Allergy, vol. 67, pp. 609–611, December 1991.
A. Miadonna, C. Zanussi, "IgE synthesis in aging. En-hancement of IgE production in subjects with autoantibo-dies", Boll. Ist. Sieroter. Milan, vol. 58, pp. 75–80, March 1979.
K. J. Erb, "Helminths, allergic disorders and IgE-mediated immune responses: where do we J. Duarte, P. Deshpande, V. Guiyedi et al., "Total and functional parasite specific IgE responses in Plasmodium falciparum-infected patients exhibiting different clinical status", Malar. J., vol. 6, pp. 1–13, January 2007.
D. Isaacs, D. G. Altman, C. E. Tidmarsh et al., "Serum immunoglobulin concentrations in preschool children measured by laser nephelometry: reference ranges for IgG, IgA, IgM", J. Clin. Pathol., vol. 36 (10), pp. 1193–1196, October 1983.
J. W. Skoug, H. L. Pardue, "Effects of reaction variables on nephelometric and turbidimetric responses for the im-munochemical reaction of immunoglobulin G", Clin. Chem., vol. 34 (2), pp. 300-308, February 1988.
V. S. Chowdary, E. C. Vinaykumar, J. J. Rao et al., "A study on serum IgE and eosinophils in respiratory allergy patients", Indian J. Allergy Asthma Immunol., vol 17 (1), pp. 21–24, January 2003.
B. Lanier, "Anti-IgE as a practical tool in asthma man-agement: observations and warnings from clinical expe-rience", Cur. Allergy Clin. Immunol., vol. 18 (3), pp. 104-106, August 2005.
N. Novak, T. Bieber, "Allergic and non-allergic forms of atopic disease", J. Allergy. Clin. Immunol., vol. 112, pp. 252–262, January 2003.
A. V. Kulakov, S. V. Klimov, D. A. Yarilin et al., "The study of the natural affinity of the human serum antibodies to component of the bacterial cell wall – muramyl dipeptide glycoside P. Tijssen, "Practice and theory of enzyme immunoassays", Lab. Techiques in Biochem. and Molecular Biology, vol. 15, pp. 558-562, 1985.
O. Yu. Galkin, A. A. Savchenko, K. I. Nikitina et al., "Ob-taining and study of the properties of new monoclonal an-tibodies to human IgE", Ukrainian Biochem. J., 2013. (in press). (in Ukrainian).
V. P. Shirobokov, I. V. Nikolaenko, L.V. Kopanytsya et al., "The panel of monoclonal antibodies for inside type diffe-rentiation of polioviruses type II", vol. 59 (6), pp. 27-36, Microbiol. J., December 1997. (in Ukrainian).