Influence of the size of the polar head of non-ionic detergents on membrane proteins immunoaffinity purification.
Nonionic polyoxyethylene type detergents (CxEy) are widely used to solubilize and purify membrane proteins. The detergent hydrophobic moiety (Cx) replaces phospholipids at exposed hydrophobic regions of the membrane proteins. During chromatography on an immobilized anti-Kell antibody to purify Kell protein (an integral erythrocyte protein), it was observed that the size of the polar head of an non ionic detergent added to the mobile phase appeared to influence the interaction of the detergent-protein complex with the immobilized antibody. Further studies were performed using another erythrocyte membrane protein, Glycophorin C and three anti-GPC monoclonal antibodies directed against three epitopes of the extracytoplasmic domain of the protein. The interaction of GPC with the three Protein A-coupled monoclonal antibodies was studied in the presence of three detergents C12E<9>, C13E<15> and C12E<23>. It was observed in batch mode and in column chromatography experiments that the adsorption of GPC to the immunoaffinity supports decreased as the size of the detergent polar head increased. Thus, the polyoxyethylene chain of a detergent might prevent the interaction of the detergent-protein complex with the immobilized antibody.
Khamlichi
S
S
INSERM U 76 INTS, Paris, France.
Loirat
M J
MJ
Blanchard
D
D
le Maire
M
M
Bailly
P
P
Cartron
J P
JP
Bertrand
O
O
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004910:Erythrocyte Membrane; D006801:Humans; D007626:Kell Blood-Group System / Q000302:isolation & purification*; D008565:Membrane Proteins / Q000302:isolation & purification*; D051379:Mice / Q000276:immunology; D013329:Structure-Activity Relationship
D000911:Antibodies, Monoclonal; D003902:Detergents; D007626:Kell Blood-Group System; D008565:Membrane Proteins