Repeated cleaning of protein A affinity column with sodium hydroxide.
Protein A attached to an affinity adsorbent was shown to be remarkably resistant to 0.5 M NaOH. Even repeated treatments gave only a small decrease in functional capacity and no adverse effect on leakage of the protein A into the eluate. This simple cleaning procedure should be useful in applications where antibodies need to be purified free from the risk of contamination with endotoxins or micro-organisms, e.g., for in vivo therapy, either in clinical trials or experimental animals. It can also prevent cross-contamination when the same protein A adsorbent is used for different batches of antibody.
Hale
G
G
Department of Pathology, University of Cambridge, UK.
Drumm
A
A
Harrison
P
P
Phillips
J
J
D000327:Adsorption; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity / Q000379:methods; D003902:Detergents; D005240:Feasibility Studies; D006801:Humans; D051379:Mice; D051381:Rats; D012972:Sodium Hydroxide; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000302:isolation & purification*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D003902:Detergents; D013205:Staphylococcal Protein A; D012972:Sodium Hydroxide