METHOD FOR RECOVERING AND PURIFYING TROPHOBLASTIC BETA-1-GLYCOPROTEIN Russian patent published in 2009 - IPC A61K35/16 A61K38/17 

FIELD: medicine.

SUBSTANCE: method for recovering and purifying trophoblastic beta-1-glycoprotein (TBG) from retroplacental blood serum by ammonium sulphate fractionation, washing and centrifugation of the solution residual matter, affinity sepharose chromatography followed by elution and lyophilisation of an end product. The retroplacental blood is centrifugated at 10000 rpm within one hour. Sodium chloride is added to a supernatant to end concentration 0.1-1.0 mol/l, while triton X-100 is added to end concentration 0.01-0.9%. Centrifugation is performed once again. The prepared serum is saturated with ammonium sulphate to 30%, incubated within a night in stirring, centrifugated. The deposit is threefold washed in 30% ammonium sulphate, suspended in 0.15M phosphate buffered saline pH 7.0-7.5, containing 0.5 M sodium chloride and 0.05% triton X-100, dialysed in an ultrafiltration cell, with tenfold amount of the buffered saline passed, five-fold diluted and ultrafiltered at filtration limit 100000 D. The prepared permeate is processed with single affinity sepharose chromatography with immobilised monoclonal TBG antibodies. The affinity-bound TBG is eluted with 0.1M glycine-Hcl buffer solution pH 2.55. The eluate is immediately neutralised, concentrated, dialysed against physiological saline and lyophilised.

EFFECT: higher yield and purity of the end product.

2 ex, 1 tbl