A simplified method for the preparation of WGA-Sepharose 4B and its use in the purification of MN blood group antigens.
A simplified method for the preparation of wheat germ agglutinin(WGA)-Sepharose 4B by coupling highly purified WGA, prepared by improved affinity chromatography, with BrCN activated Sepharose 4B in a solution of high carbonate buffer is described. The amount of WGA linked to Sepharose 4B was 82.40% (3.07 mg WGA per ml Sepharose 4B). MN blood group antigens of human erythrocyte membranes purified with WGA-Sepharose 4B affinity chromatography showed a single band on sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS/PAGE). The yield of the antigens from 400 mL fresh blood was 32-40 mg. The WGA-Sepharose 4B column could be used several times without loss of activity.
Zhang
S M
SM
Department of Biochemistry, Second Military Medical College, Shanghai, People's Republic of China.
Li
J X
JX
Wei
Y M
YM
Yang
Z M
ZM
D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007519:Isoantigens / Q000302:isolation & purification*; D008951:MNSs Blood-Group System / Q000276:immunology*; D012685:Sepharose / Q000031:analogs & derivatives
D007519:Isoantigens; D008951:MNSs Blood-Group System; C042758:wheat germ agglutinin-sepharose 6MB; D012685:Sepharose