BIOLOGICAL ASSAY OF PEPTIDOGLYCANS Russian patent published in 2019 - IPC G01N33/50 

FIELD: chemistry.

SUBSTANCE: method includes a) treating a glucose polymer sample with ultrasound, heating and/or alkalising to fragment and break down the PGN contained in the sample and to form soluble PGN weighing between 30 and 5,000 kDa; b) bringing the treated sample into contact with a recombinant cell expressing an exogenous TLR2 receptor (Toll-like receptor 2) and the reporter gene, with direct dependence on the signalling pathway associated with the TLR2 receptor, the reporter gene encoding secreted alkaline phosphatase; c) measuring the reporter gene signal and d) determining the amount of PGN in the sample using a calibration curve based on the relationship between the amount of PGN and the intensity of the reporter gene signal, where the calibration curve of the amount of PGN versus the intensity of the reporter gene signal is standardised or calibrated using PAM₃Cys-Ser-(Lys)4 trichlorohydrate. Also disclosed is a kit for assay of peptidoglycans (PGN) in a sample of glucose polymers. Kit contains PAM3Cys-Ser-(Lys)4 trichlorohydrate; a recombinant cell expressing exogenous TLR2 receptor and reporter gene with direct dependence on the signalling pathway associated with the TLR2 receptor, the reporter gene encoding secreted alkaline phosphatase; and either a calibration curve based on the amount of PGN as a function of the intensity of the reporter gene signal, either a PGN standard, preferably derived from a bacterium selected from Staphylococcus aureus, Micrococcus luteus, Bacillus subtilis and Alicyclobacillus acidocaldarius, preferably from Staphylococcus aureus, Micrococcus luteus and Alicyclobacillus acidocaldarius; optional instructions for use and a solution for pre-treatment of the sample.

EFFECT: invention relates to a method for assay of peptidoglycans (PGN) in a glucose polymer sample.

10 cl, 8 dwg