Patent GB-779493-A (GB)
Title: Process for the isolation of stable, highly purified gamma globulin preparations
IPC: C07K16/06
Filed: 1953-08-04
Abstract: Stable highly purified gamma globulin preparations of uniform molecular size are prepared from human or animal blood serum or plasma containing antibodies by precipitating the main portion of the beta globulins without the use of protein denaturating solvents from a crude albumin-free globulin fraction at an ionic strength of 0.07\sB0.02, at a pH of 5.3\sB0.05 and a protein concentration of about 3-5 per cent, the main portion of alpha globulins being then precipitated at an ionic strength below about 0.001, at a pH of 5.0\sB0.05 and a protein content of 2\sB0.5 per cent, and the residual quantities of alpha and beta globulins are separated as insoluble aluminium compounds by addition of aluminium hydroxide gel at a pH of 5.0\sB0.05.  The procedure is detailed in the examples. In Example 1, human blood serum is diluted with water to a protein content of 3.8 per cent, the pH adjusted to 8.0 and the globulins precipitated by addition of saturated ammonium sulphate.  The precipitate is dissolved in water, freed from traces of sediment and dialysed.  The ionic strength is adjusted to 0.07 and the solution acidified to pH 5.3\sB 0.05 by addition of acetic acid to precipitate the beta globulins which are removed by centrifuging.  The solution is dialysed to an ionic strength of below 0.001, further diluted to a protein content of 2 per cent and acidified to pH 5.0\sB0.05 by addition of acetic acid.  The precipitate is removed and further residual alpha and beta globulins in the solution adsorbed on aluminium hydroxide gel.  The purified solution may be lyophilized. Specification 669,461 and U.S.A.  Specifications 2,437,060 and 2,543,215 are referred to.ALSO:Stable highly purified gamma globulin preparations of uniform molecular size are prepared from human or animal blood serum or plasma containing antibodies by precipitating the main portion of the beta-globulins without the use of protein denaturating solvents from a crude albumin-free globulin fraction at an ionic strength of 0.07 \sB 0.02, at a pH of 5.3 \sB 0.05 and a protein concentration of about 3-5 per cent., the main portion of alpha globulins being then precipitatd at an ionic strength below about 0.001, at a pH of 5.0 \sB 0.05 and a protein content of 2 \sB 0.5 per cent and the residual quantities of alpha and beta globulins are separated as insoluble aluminium compounds by addition of aluminium hydroxide gel at a pH of 5.0 \sB 0.05.  The procedure is detailed in the examples.  In Example 1, human blood serum is diluted with water to a protein content of 3.8 per cent. the pH adjusted to 8.0 and the globulins precipitated by addition of saturated ammonium sulphate.  The precipitate is dissolved in water, freed from traces of sediment and dialysed.  The ionic strength is adjusted to 0.07 and the solution acidified to pH 5.3 \sB 0.05 by addition of acetic acid to precipitate the beta globulins, which are removed by centrifuging. The solution is dialysed to an ionic strength of below 0.001, further diluted to a protein content of 2 per cent and acidified to pH 5.0 \sB 0.05 by addition of acetic acid.  The precipitate is removed and further residual alpha and beta globulins in the solution adsorbed on aluminium hydroxide gel.  The purified solution may be lyophilized and the purified product dissolved in physiological saline and injected.  Specification 669,461 and U.S.A. Specifications 2,437,060, 2,543,215 are referred to.