Alexandr E Urusov obtained his Doctor degree in Chemistry in A.N. Bach Institute of Biochemistry in 2012. Now he is Researcher in Research Center of Biotechnology of the Russian Academy of Sciences. His research interests molecular immunology, immunology, methods and apparatus in immunology, biotechnology, immunoenzyme analysis, medicine, health care and medical biochemistry.
Mycotoxins are one of the priority classes of agricultural products contaminants. They pose a significant danger to human and animal health. For productive monitoring of these toxic contaminants new formats of immunochromatographic (ICA) and immunoenzyme assays (IEA) have been developed. For highly sensitive detection in indirect labeling ICA, the combination of non-modified specific antibodies and colloidal gold nanoparticles-labeled anti-species antibodies (anti-IgG-GNP) is proposed. This approach allows decreasing limits of detection in 10-20 times (for example, for zearalenone to 350 pg/ml). The assay duration varies from 10 to 20 min. For IEA, the use of immunosorbents based on magnetic nanoparticles is proposed. The given approach allows concentrating the analyte and performing immunochemical interactions in homogeneous, diffusion-independent mode, thereby reducing the detection limit and the duration of the analysis. The total IEA duration is reduced by 3-4 times - up to 20-30 min (with 5-min specific immune interaction). The limit of detection is decreased on an order of magnitude as compared with common heterogeneous microplate IEA, accounting for example, 2 pg/ml for aflatoxin B1. The developed assays for aflatoxin B1, ochratoxin A, zearalenone and T2-toxin ensure 90% or higher revealing the analyte in cereals, thus indicating prospects of the proposed approaches.
Zvereva E A has graduated from the Moscow Institute of Food Production and works in A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences as a Senior Researcher. Her scientific interests are in the field of development of new immunochemical techniques for the detection of biologically active compounds - pesticides, hormones and antibiotics. These works cover the full cycle of the development of immunoanalytical techniques, beginning from obtaining immune reactants and finalizing by the creation of manufacturing technologies.
Statement of the Problem: Cow\'s milk and products derived from it are an essential component of a food ration for many people and a source of a number of valuable biologically active compounds. However, at the same time, milk is one of the most common food allergens. Different processing technologies are actively developed to reduce the allergenicity of dairy products while preserving their nutritional value and functional properties. In this connection, analytical methods are required for correct assessment of the content of allergenic components in dairy products. Immunochemical methods provide such characteristic with high selectivity and productivity. However, the choice of their formats to work with wide variety of food stuffs requires additional studies.\r\n\r\nMethodology: The study was focused on immunodetection of such main allergenic compounds of milk and dairy products as β-lactoglobulin (BLG), α-lactalbumin (ALA) and bovine serum albumin (BSA). Enzyme-linked immunosorbent assays (ELISA) have been developed and characterized for this purpose.\r\n\r\nFindings: Concentration and kinetic dependences of the analytical interactions were studied and the optimal modes of immunodetection were determined. The chosen protocols allowed carrying out all ELISA stages at room temperature. Advantages of competitive assay format for the detection of allergenic structures in partially hydrolyzed proteins have been confirmed in comparison with sandwich assay format. The developed ELISAs were characterized by detection limits of 10, 4.5, and 13.4 ng/mL for BLG, ALA and BSA, respectively. The BLG ELISA was used for testing milk and dairy products, including products with reduced allergenicity that were obtained by enzymatic hydrolysis of raw milk. \r\n\r\nConclusion & Significance: The developed methods allow controlling both intermediate technological products in dairy industry and final food stuffs. Their use will provide efficient protection of sensitive consumers from milk allergens.\r\n