Pilot scale purification of human monoclonal IgM (COU-1) for clinical trials

Ida Tornøe, Ingrid L. Titlestad*, Karin Kejling, Karin Erb, Henrik J. Ditzel, Jens C. Jensenius

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review


No standard procedure is available for the purification of human monoclonal antibodies for human i.v. administration. Here we describe the procedure developed for pilot scale purification of the human IgM monoclonal antibody COU-1 directed against a cancer-associated antigen. The hybridoma cells were grown in protein-free medium and purification from the clarified culture supernatant was carried out in 4 simple chromatographic steps: (1) hydroxylapatite chromatography: (2) hydrophobic interaction chromatography on phenyl-Sepharose: (3) cation-exchange chromatography on sulphonyl-Sepharose; and (4) anion-exchange chromatography on tetraethylamino-Sepharose. The product was substantially pure with regard to protein after step 3, but contained DNA which was removed in step 4. The average recovery of the IgM was 54% with a range of 40-65%. Importantly, the ability of the antibody to bind to its antigen in ELISA was fully maintained during the purification. Subsequently, the purified antibody was isotope labelled and successfully used for in vivo detection of colon, rectal and pancreas carcinomas in patients. The purification procedure described appears to compare favourably with previously published methods, but a critical comparison is not possible due to the lack of necessary information in the available literature.

Original languageEnglish
JournalJournal of Immunological Methods
Issue number1
Pages (from-to)11-17
Number of pages7
Publication statusPublished - 23. Jun 1997


  • Human monoclonal antibody
  • IgM purification
  • Pilot scale


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