Fractionation of IgG, IgG2a, IgG2b and IgG3 immunoglobulins from mouse serum by protein A-sepharose column chromatography.

摘要: Normal mouse serum was fractionated on protein A-Sepharose column by the elution with citrate buffer of different pH. At pH 8.0 IgG all subclasses were retained and main portion IgG1, IgG2a, IgG3, IgG2b eluted at 6.0, 5.0, 4.0 3.0, respectively, as assessed Ouchterlony immunodiffusion. Pure IgG1 obtained 6.0 from normal serum. The identity globulin confirmed its antibody activity in chromatography fraction anti-sheep red blood cell antiserum also inability to fix complement. IgG3 5.0. formation M-bow-like precipitin line gamma region immuno-electrophoresis. affinity A reduced treatment 2-mercaptoethanol (2-ME). From 2-ME treated serum, therefore, pure IgG2a could be 4.0, although recovery low. Otherwise, always other subclass, or IgG3.