Development of Enzyme-Linked Immunoassay Systems for Monitoring the Content of the Main Non-Meat Additives in Meat Products

Abstract

Abstract-Methods for control of the content of non-meat components (connective tissue of animals, eggs, soybeans) in meat products have been developed based on competitive enzyme immunoassay of biomarker proteins: collagen, ovalbumin, and soybean trypsin inhibitor. Polyclonal rabbit antibodies against the biomarkers were produced. Screening of the following analysis conditions was carried out using the most affine preparations: the duration of the stages, the concentrations of the reagents, the composition of the reaction medium to ensure the completeness and the minimum limits of detection of analytes. It was shown that the immunochemical stage can be transferred to the kinetic mode and reduced to 20 min, while the total analysis took only 1 h. The selected conditions synchronized detection stages and provided the same signal amplitudes from all three biomarkers. An additional advantage of the method is that the analysis can be carried out at room temperature. The detection limits for collagen, ovalbumin and soybean trypsin inhibitor in the final extracts were 0.025 μg/mL, 0.012 μg/mL, and 0.001 μg/mL, respectively. Approbation of the method showed the reliability of the conclusions about the composition of the tested meat products; the relative standard deviation in the analysis of the monitored biomarkers was 8-10%. Key words: enzyme-linked immunoassay, meat products, collagen, ovalbumin, soybean trypsin inhibitor The work was financially supported by the Russian Science Foundation (project no. 19-16-00108).