A rapid method for purification of monoclonal human IgM from mass culture

Article type: Research Article

Authors: Vollmers, H. Peter^{a; *} | Wozniak, Ewa^a | Stepien-Bötsch, Ewa^a | Zimmermann, Ulrich^b | Müller-Hermelink, Hans-Konrad^a

Affiliations: [a] Institut für Pathologie, Universität Würzburg, Josef-Schneider Würzburg, Germany | [b] Institut für Biotechnologie, Universität Würzburg, Am Hubland, Würzburg, Germany

Correspondence: [*] Correspondence and reprint requests to: Dr H. Peter Vollmers, Institut für Pathologie, Universität Würzburg, Josef-Schneider Str. 2, 97080 Würzburg, Germany.

Abstract: 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 RPM1 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.

Keywords: Human immunoglobulin M, immunoglobulin mass culture, water dialysis, pure human IgM

DOI: 10.3233/HAB-1996-7105

Journal: Human Antibodies, vol. 7, no. 1, pp. 37-41, 1996