High quality human immunoglobulin G purified from Cohn fractions by liquid chromatography

ABSTRACT

In order to obtain intravenous immunoglobulin G (iv IgG) of high quality from F-I+II+III or F-II+III pastes prepared by the Cohn method, we developed a chromatography process using ion exchange gels, Q-Sepharose FF and CM-Sepharose FF, and Sephacryl S-300 gel filtration. Viral inactivation was performed by incubating the preparation with pepsin at pH 4.0 at 35oC for 18 h. The characteristics of 28 batches produced by us were yield 4.3 + or - 0.2 g/l plasma, i.e., a recovery of 39.1 + or - 1.8 percent; IgG subclasses distribution IgG1 = 58.4 percent, IgG2 = 34.8 percent, IgG3 = 4.5 percent and IgG4 = 2.3 percent; IgG size distribution was 98.4 percent monomers, 1.2 percent dimers and 0.4 percent polymers and protein aggregates; anticomplement activity was less than 0.5 CH50/mg IgG, and prekallikrein activator activity (PKA) was less than 5 IU/ml. These characteristics satisfied the requirements of the European Pharmacopoea edition, and the regulations of the Brazilian Health Ministry (M.S. Portaria No. 2, 30/10/1998)