Process for the industrial-scale purification of gamma globulins from human plasma for industrial applications

摘要

The invention relates to a novel, industrial-scale process for the purification of gamma-immunoglobulins (IgG) starting from plasma or fractions thereof. The method involves two chromatographic steps, i.e. a cation exchange capture chromatography, and then a polishing anion exchange chromatography, ensuring a highly purified end product, which contains no aggregates, and high yields. The process also involves a virus inactivation step by means of a solvent/detergent treatment to inactivate the viruses with a lipid envelope, and a virus removal step by nanofiltering to ensure the removal of the non-enveloped viruses.

主权项

1. A process for the production of IVIG, employing as starting material human plasma or an intermediate plasma fraction enriched in IgG, said process comprising the following chromatographic steps: a first chromatographic step, consisting in a step of “capture” of IgG, performed on a strong cation exchanger and a second chromatographic step, consisting in a step of “polishing”, performed on a strong anion exchanger;
wherein the strong cation exchanger belongs to the category of strong exchanger characterized by SO3−functional group; and wherein the strong anion exchanger has quaternary ammonium groups as exchanger groups; wherein between the two chromatographic steps is performed at least a step of viral inactivation performed by means of a solvent-detergent treatment; wherein subsequently after the second chromatographic step the solution containing IVIG is subjected to a viral inactivation step by means of nanofiltration; wherein strong cation exchanger is charged with 20-60 mg of IgG/ml of resin and wherein the conditioning, charging and washing steps are performed employing a 25-75 mM buffer at pH 5.4-5.7 while the elution is performed using a buffer having 0.25-1 M sodium chloride concentration; wherein the strong anion exchanger is charged with 20-60 mg of protein/ml of resin and wherein the conditioning, charging and washing steps are performed employing a 10-30 mM buffer, pH 9.0±0.5, NaCl 8-10 mM while the elution is performed or using an increased ionic strength or by means of pH variation; when the elution is performed using an increased ionic strength it is employed a buffer having a conductibility of 9.5-10.5 mS/cm and pH 9.0±0.5; when the elution is performed by means of pH variation, it is employed a buffer having a a conductibility of 1-1.5 mS/cm, and pH 6-7.