Purification of murine IgG3 and IgM monoclonal antibodies by euglobulin precipitation.
We describe a simple and efficient non-chromatographic method for the purification of murine IgG3 and IgM monoclonal antibodies (MAbs) which takes advantage of their euglobulin properties. Following filtration, ascitic fluid is dialysed against demineralized water and centrifuged at 22,000 X g for 30 min. The resulting precipitate is dissolved in a high salt buffer (0.1 M Tris-HCl, 1 M NaCl, pH 8). A second cycle of dialysis and centrifugation yields a product of high purity. Nine IgG3 MAbs and eight IgM MAbs were purified by this procedure. Recovery was greater than 90% for seven of nine IgG3 MAbs. It was less reproducible for IgM MAbs and ranged from 40% to greater than 90% depending on antibody and batch. Purity was assessed by SDS-polyacrylamide gel electrophoresis. The purified immunoglobulin was essentially free of albumin, transferrin, and other mouse ascites proteins. No loss of antibody function was observed.
García-González
M
M
Clonatec, Paris, France.
Bettinger
S
S
Ott
S
S
Olivier
P
P
Kadouche
J
J
Pouletty
P
P
D000818:Animals; D000907:Antibodies, Bacterial / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D000913:Antibodies, Protozoan / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000302:isolation & purification; D000937:Antigen-Antibody Reactions; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000302:isolation & purification*; D007075:Immunoglobulin M / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D011233:Precipitin Tests / Q000379:methods*; D012712:Serum Globulins / Q000276:immunology*
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D000914:Antibodies, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D012712:Serum Globulins