ES398015A1 - Process for isolating a fibrin-stabilizing factor - Google Patents

Abstract

Procedure for the isolation of a fibrin stabilizing factor, characterized in that, in the cold, a) an extract released from solid impurities, prepared by means of NaCl solution from human placentas, is mixed at pH 5.0-7.5, preferably 6.0, with a solution of diaminoethoxy acridinlactate; b) the separated precipitate, optionally after several washes, is collected in dilute alkali metal chloride solution, preferably NaCl solution, of pH 7.0-8.0, preferably 7.5, containing approximately 5% of a complex forming agent (based on alkali metal chloride); c) from the solution, after removal of insoluble concomitant substances and optionally after further dilution, the inactive concomitant substances are precipitated by means of the addition of a quaternary ammonium base; d) from the filtrate another precipitate is obtained by diaminoethoxyacridinlactate; e) the precipitate, after removal of the supernatant portion by the addition of a dilute solution of alkali metal chloride, preferably NaCl, pH 7.0-8.0 preferably 7.5, containing approximately 5% forming agent of complexes (referred to the alkali metal chloride), is decomposed, and the residue is separated; f) the filtrate is mixed with 20-30%, preferably 25%, of solid ammonium sulfate, the precipitate is separated after a few hours, mixed with stirring with dilute solution of complexing agent to form a paste, and it is dialyzed against tris (hydroxymethyl) aminomethane and hydrochloric acid buffer, which also contains complexing agent and NaN3; g) dialyzed product that remains as residue, after removal of impurities at pH 6.0, is gel filtered at neutral pH, a precipitate is precipitated from the active fractions by solid ammonium sulfate, and dissolved in the precipitate in neutral buffer of tris- (hydroxymethyl) -aminomethane and ethylenediaminetetraacetic acid; h) the solution is dialyzed against neutral buffer of tris (hydroxymethyl) -aminomethane and ethylenediaminetetraacetic acid, from the remaining solution factor XIII is precipitated at pH approximately 5.0 as euglobulin; i) the separated precipitate is dissolved in physiological NaCl solution, which contains a small amount of complexing agent, the solution is mixed with a stabilizer, filtered in sterile medium, dialyzed, normalized and optionally lyophilized. (Machine-translation by Google Translate, not legally binding)