METHOD FOR DETERMINATION FUNCTIONAL ACTIVITY OF C3-CONVERTASE OF ALTERNATIVE PATH OF HUMAN COMPLEMENT ACTIVATION Russian patent published in 2018 - IPC G01N33/50 

FIELD: immunology.

SUBSTANCE: rabbit erythrocyte lysis reaction is carried out with human serum in VBS-Mg²⁺-EGTA conditions at a temperature of 37.0±0.5 °C with the subsequent determination of the degree of lysis of erythrocytes spectrophotometrically at a wavelength of 405 nm. In this case, the lysis of the rabbit's erythrocytes is carried out with 3.5 % of human blood serum for 10 minutes, the complement activation reaction is stopped by the addition of 100 mcl of veronal buffer containing 10 mM EDTA, 2.4 ml of a chilled solution is added to the control sample 0.15 M NaCl, centrifuge and the determine erythrocyte lysis degree [Y(control), %], reincubate the test sample for 30 min at 37 °C, determine the degree of lysis of erythrocytes [Y(experiment), %]. Then, the functional activity of the C3 convertase of the alternative pathway of activation of the complement system (APAC) is calculated as the difference between the degree of erythrocyte lysis in the test and control assays and with a difference in the degree of lysis between the test and control samples to 15 % take as normal size the functional activity of the C3-convertase of the alternative complement pathway in the blood serum of the patient.

EFFECT: method for determining the functional activity of the C3-convertase of APAC is proposed.

1 cl, 1 dwg, 4 tbl, 1 ex