A rapid method for purification of monoclonal human IgM from mass culture.
We describe in this paper the rapid and cheap purification of human immunoglobulin M from hybridoma supernatant of mass culture. The method consists of two steps: 1. concentration of supernatant by ultrafiltration and 2. dialysis against distilled water, pH 6.4. To produce the supernatant, the hybridomas are grown in RPMI media with fetal calf serum (10% FCS) in 250 ml flasks under normal tissue culture conditions. More than 14 mg IgM can be nearly selectively purified within 6 h from 5 l of antibody containing hybridoma supernatant. Most of the IgM molecules stay in a penta- and monomeric form and are positive in physiological and immunohistochemical studies.
Vollmers
H P
HP
Institut für Pathologie, Universität Würzburg, Germany.
Wozniak
E
E
Stepien-Bötsch
E
E
Zimmermann
U
U
Müller-Hermelink
H K
HK
D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D003956:Dialysis / Q000379:methods*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007075:Immunoglobulin M / Q000302:isolation & purification*; D009101:Multiple Myeloma / Q000276:immunology; D013154:Spleen / Q000166:cytology / Q000276:immunology; D013274:Stomach Neoplasms / Q000276:immunology; D014867:Water
D000911:Antibodies, Monoclonal; D007075:Immunoglobulin M; D014867:Water