Purification of mouse monoclonal antibodies by chromatography on sepharose 6 B.
By using Sepharose 6 B, a simple procedure for purification of mouse monoclonal antibodies (mcAbs) of the IgM and IgG class from ascites has been developed. The procedure which was applied to purify mcAbs against insulin and pancreatic islet cells permits either direct chromatographic separation from ascites protein components or after precipitating the immunoglobulins with ammonium sulphate. Recovery of the immunoglobulins was found to be approximately 80%, and the immunological reactivity, as tested by indirect immunofluorescence and ligand binding assay, was almost completely retained. For purification of IgG from ascites, precipitation with ammonium sulphate is recommended prior to chromatography on Sepharose 6 B, whereas IgM can directly be subjected without any pretreatment.
Diaz-Alonso
J M
JM
Central Institute of Diabetes, Gerhardt Katsch, Karlsburg.
Kohnert
K D
KD
Keilacker
H
H
Ziegler
B
B
Ziegler
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001202:Ascitic Fluid / Q000032:analysis; D002850:Chromatography, Gel; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification*; D007075:Immunoglobulin M / Q000302:isolation & purification*; D007328:Insulin / Q000276:immunology; D007515:Islets of Langerhans / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D012685:Sepharose
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007328:Insulin; D012685:Sepharose