Patent GB-731104-A (GB)
Title: Improvement in preparation of blood fraction for use in rh testing procedure
IPC: G01N33/80
Filed: 1953-02-27
Abstract: A serum fraction for use in Rh testing procedures is produced by passing bovine blood serum through a column containing both cation and anion exchange resins in a proportion such that a solution having a pH of 6 to 8, and preferably 7.0, results on passage through the column of an acidic, basic or neutral serum, whereby a precipitate of euglobulins is formed in the solution. The precipitate is removed and to the resultant solution is added an alkali metal salt of a fatty acid having from 4 to 10 carbon atoms, or an alkali metal salt of acetyltryptophane, in an amount sufficient to bring the molarity of the solution with respect thereto within the range of from 0.15 to 0.3.  The pH is adjusted to 6.8 to 7.4 and the solution heated to 75 DEG  C. for 25 to 35 minutes.  After cooling to 25 DEG  C., the pH is adjusted to 5.5 to 5.8 and the precipitated denatured globulins are removed.  The alkali metal salt of the fatty acid or acetyltryptophane is removed by, e.g., dialysis or ion exchange resins and the resultant solution, consisting of albumin and alpha-globulins, is adjusted to pH 7.0 and concentrated or dried. Alternatively, after removing the precipitated denatured globulins, the solution may be adjusted to pH 3.0 to 5.5, and preferably 4.4 to 4.8, to precipitate the alpha-globulins, cooled to 2 DEG  to -1 DEG C. and chilled methanol added to at least 38 per cent by volume to precipitate the albumin, the alpha-globulins and albumin then being separated from the solution.  The test solution contains 26 to 30 per cent of the albumin and alpha-globulin proteins in normal saline and has a pH of 7.0 to 7.4.  The cation-exchange resins are such as have active phenolic, carboxylic or sulphonic groups which are saturated with hydrogen ions, e.g. phenol methylene sulphonic acid resin, nuclear sulphonic acid resin and polymerized acrylic acid resin.  The anion exchange resins have active amine groups such as a polymer resulting from nitration and subsequent reduction of a styrene divinylbenzene copolymer.ALSO:A serum fraction for use in Rh testing procedures is produced by passing bovine blood serum through a column containing both cation and anion exchange resins in a proportion such that a solution having a pH of 6 to 8, and preferably 7.0, results on passage through the column of an acidic, basic or neutral serum, whereby a precipitate of euglobulins is formed in the solution. The precipitate is removed and to the resultant solution is added an alkali metal salt of a fatty acid having from 4 to 10 carbon atoms, or an alkali metal salt of acetyltryptophane, in an amount sufficient to bring the molarity of the solution with respect thereto within the range of from 0.15 to 0.5.  The pH is adjusted to 6.8 to 7.4 and the solution heated to 75 DEG C. for 25 to 35 minutes.  After cooling to 25 DEG C., the pH is adjusted to 5.5 to 5.8 and the precipitated denatured globulins are removed.  The alkali metal salt of the fatty acid or acetyltryptophane is removed by e.g. dialysis or ion exchange resins and the resultant solution, consisting of albumin and alpha-globulins, is adjusted to pH7.0 and concentrated or dried. Alternatively, after removing the precipitated denatured globulins, the solution may be adjusted to pH 3.0 to 5.5, and preferably 4.4 to 4.8 to precipitate the alpha-globulins, cooled to 2 DEG  to -1 DEG C. and chilled methanol added to at least 38 per cent by volume to precipitate the albumin, the alpha globulins and albumin then being separated from the solution.  The test solution contains 26 to 30 per cent of the albumin and alpha globulin proteins in normal saline and has a pH of 7.0 to 7.4.  The cation exchange resin are such as have active phenolic, carboxylic or sulphonic groups which are saturated with hydrogen ions e.g. phenol methylene sulphonic acid resin, nuclear sulphonic acid resin and polymerised acrylic acid resin.  The anion exchange resins have active amine groups such as a polymer resulting from nitration and subsequent reduction of a styrene divinylbenzene copolymer.