Scientific journal
Scientific Review. Biological science
ISSN 2500-3399
ПИ №ФС77-57454

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DEVELOPMENT OF METHOD FOR DETERMINATION OF BETA-LACTOGLOBULIN IN MILK AND MILK PRODUCTS BY ENZYME-LINKED IMMUNOSORBENT ASSAY

Zvereva E.A. 1 Smirnova N.I. 1 Zherdev A.V. 1 Dzantiev B.B. 1 Yurova E.A. 2 Denisovich E.Y. 2 Zhizhin N.A. 2 Kharitonov V.D. 2 Agarkova E.Y. 2 Botina S.G. 2 Ponomareva N.V. 3 Melnikova E.A. 4
1 A.N. Bach Institute of biochemistry
2 All-Russian research institute of dairy industry
3 Voronezh state university of engineering technologies
4 Public corporation Dairy combine “Voronezhsky”
Method for determination of beta-lactoglobulin (BLG) in milk and milk products by enzyme-linked immunosorbent assay (ELISA) has been developed. This method is used for the content control of beta-lactoglobulin in milk and milk products, including products with reduced allergenicity, obtained by enzymatic hydrolysis of milk protein. Concentration and kinetic characteristics of the interactions in the immunoassay system were studied and the optimal mode of immunodetection has been determined. It is shown that the optimal analytical parameters are provided when BLG is immobilized in ELISA microplate wells from a concentration of 5 ng/ml and the antibodies against BLG are used in a concentration of 0.2 ug/ml. Shortening competitive step twice (from conventional 1 h to 30 min) does not lead to loss neither sensitivity nor amplitude of the detected signal. The proposed method allows to carry out all stages of the analysis at room temperature. Working range of the measured BLG concentration is from 0.03 to 2.5 ug/ml. The coefficient of variation in the working range is from 1.8 % to 9.2 %; the reproducibility of measurements in the course of a week - not more than 10.5 %.