Separation of ovine IgG1 and IgG2 on protein A-sepharose.
A method using protein A-Sepharose chromatography was developed to separate and purify ovine IgG1 and IgG2. The IgG1 eluted from protein A-Sepharose at pH 6.8 and IgG2 eluted at pH 4.5. This method was used to show the specific transfer of IgG1 from the colostrum to newborn lambs. After separation on protein A-Sepharose both IgG1 and IgG2 were pure as analyzed by isoelectric focusing, Western Blotting and SDS-PAGE. The isoelectric points for the immunoglobulins were calculated to be 3.5 for IgG2 and a range from 6.2 to 8.1 for IgG1. The subclass, IgG1, was present in the whey and was the subclass that was found in the serum of lambs after being fed colostrum. The ewe sera had a decrease of both IgG1 and IgG2 at the time of lambing compared to 2 weeks prior to parturition.
Schmerr
M J
MJ
National Animal Disease Center, USDA, Agricultural Research Service, Ames, IA 50010.
Goodwin
K R
KR
D000818:Animals; D015153:Blotting, Western; D002847:Chromatography, Agarose; D003126:Colostrum / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D007112:Immunity, Maternally-Acquired; D007074:Immunoglobulin G / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D012685:Sepharose; D012756:Sheep / Q000276:immunology*; D013205:Staphylococcal Protein A
D007074:Immunoglobulin G; D013205:Staphylococcal Protein A; D012685:Sepharose