METHOD OF DETERMINING THE SPECIFIC ACTIVITY OF CETUXIMAB USING A BIOLOGICAL METHOD ON CELL CULTURE Russian patent published in 2023 - IPC G01N33/15 

FIELD: immuno-oncology.

SUBSTANCE: invention relates to a method of determining the specific activity of Cetuximab by a biological method on cell culture. The A431 cell line is seeded in an amount of 10,000 cells into a 96-well plate and serial dilutions of cetuximab in the range of 7,812.5 ng/ml — 30.5 ng/ml are added in triplicate to the cell plate 4 hours after cell seeding. The A431 cell line is incubated with serial dilutions of cetuximab in a 96-well plate for 96 hours at 37°C in three repetitions. Then 10 μl of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) at a concentration of 5 mg/ml is added to each well and incubated for 4 hours. Formazan crystals are dissolved in preheated to 37°C lysis buffer with a pH of 4.7, consisting of sodium acetate, sodium lauryl sulfate, dimethylformamide and purified water. Using a shaker it is mixed for 3 hours while maintaining the speed at 450 rpm and a temperature of 37°C. Colorimetric evaluation is carried out using a plate spectrophotometer at a wavelength of 570 nm.

EFFECT: invention makes it possible to evaluate the specific activity of the medicinal product, which characterizes the pharmacological action of the medicinal product, and can be used for more effective treatment of oncological diseases by adapting treatment to a specific patient.

1 cl, 10 dwg, 1 tbl, 5 ex