A one-step purification method for monoclonal antibodies based on salt-promoted adsorption chromatography on a 'thiophilic' adsorbent.
A convenient and fast method for the purification of mouse monoclonal antibodies from the culture media of cloned cells or from ascites fluids by means of salt-promoted chromatography on a 'thiophilic' adsorbent is described. The adsorbent has a capacity to adsorb about 20 mg/ml of immunoglobulins and a broad specificity towards immunoglobulins derived from various animal species irrespective of the type or subclass to which they belong. The recovery of the purified IgG is better than 90% while that for IgM is considerably less, probably due to dissociation occurring during the adsorption-desorption process. This one-step procedure also leads to a considerable concentration of dilute solutions of immunoglobulins. Moreover, the purified Igs are eluted by an essentially salt-free buffer at near neutral pH thus obviating the need for post-treatment of the sample before storage or subsequent conjugation to enzymes for use in immunoassays. This purification method is also well suited for large-scale operations since sample preparation requires only the addition of 0.5 M K2SO4 to the ascited fluid or cell culture medium. The degree of purification obtained is, in certain instances, comparable to that obtained by biospecific affinity chromatography based on antigen-antibody interactions. In contrast to immunosorption and desorption methods, however, there is no risk of contamination of the immunoglobulins purified on the 'thiophilic' adsorbent by foreign proteins.
Belew
M
M
Institute of Biochemistry, Uppsala Biomedical Center, University of Uppsala, Sweden.
Juntti
N
N
Larsson
A
A
Porath
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D000915:Antibody Affinity; D001202:Ascitic Fluid / Q000032:analysis; D002845:Chromatography / Q000379:methods*; D003470:Culture Media / Q000032:analysis; D008623:Mercaptoethanol; D051379:Mice; D012492:Salts; D012685:Sepharose; D013045:Species Specificity; D013438:Sulfhydryl Compounds
D000911:Antibodies, Monoclonal; D003470:Culture Media; D012492:Salts; D013438:Sulfhydryl Compounds; D008623:Mercaptoethanol; D012685:Sepharose