METHOD OF DETERMINING ANTIPHOSPHOLIPID ANTIBODIES Russian patent published in 1999 - IPC

FIELD: biotechnology. SUBSTANCE: invention aims at determining antiphospholipid antibodies in the case of obstetrical complications. Phospholipid is dispersed in aqueous medium containing cryogenic protector at its weight ratio to phospholipid from 1:1 to 1:3. Phospholipid suspension is then adsorbed on polystyrene plates at 30- 40 C. Plate is treated with buffer solution at pH 7.4 ± 0.2 containing gelatin or bovine serum albumin at concentration 5-20 mg/ml. Then, plate is incubated with blood serum diluted at volume ratio 1:20 to 1:60 with above-mentioned buffer solution containing 5- 10 mg/ml of gelatin or bovine serum albumin. Plates are washed with buffer solution before and after incubation. As conjugate, that of monoclonal antibodies against human immunoglobulin with radish peroxidase is used, and, as substrate-chromogen mixture, substrate- chromogen mixture containing o-phenylenediamine and hydrogen peroxide. Optical density is measured at wavelength 492 nm. According to invention, phospholipid utilized in dispersion step is chicken yellow phosphatidylcholine or chicken yellow phosphatidylethanolamine, or bovine brain phosphatidylserine, or bovine brain sphingomyelin, or bovine heart cardiolipin, or yeast phosphatidylinositol. When dispersing phosphatidylethanolamine, acetic acid is added to aqueous medium with cryogenic protector to final concentration 10 mg/ml. EFFECT: simplified procedure and enabled reliable results on storage of prepared plates up to 6 months. 3 cl, 4 tbl, 8 ex