High quality human immunoglobulin G purified from Cohn fractions by liquid chromatography.
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 35 degrees C for 18 h. The characteristics of 28 batches produced by us were: yield 4.3 +/- 0.2 g/l plasma, i.e., a recovery of 39.1 +/- 1.8%; IgG subclasses distribution: IgG1 = 58.4%, IgG2 = 34.8%, IgG3 = 4.5% and IgG4 = 2. 3%; IgG size distribution was 98.4% monomers, 1.2% dimers and 0.4% 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).
Tanaka
K
K
Divisão de Pesquisa e Desenvolvimento Industrial, Fundação Pró-Sangue Hemocentro de São Paulo, São Paulo, SP, Brasil. georgiaandrade@originet.com.br
Sawatani
E
E
Dias
G A
GA
Shigueoka
E M
EM
Campos
T C
TC
Nakao
H C
HC
Arashiro
F
F
D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification*; D016756:Immunoglobulins, Intravenous / Q000592:standards*; D012015:Reference Standards
D007074:Immunoglobulin G; D016756:Immunoglobulins, Intravenous