title: Use of sulfonation method to determine DNA contamination of cell cultured monoclonal antibodies produced for human therapeutics.
abstract: In the past few years, culture of transformed mammalian cells has been widely used to produce natural or recombinant molecules, as monoclonal antibodies (MAb) and cytokines. For therapeutic use, the cellular DNA level must be determined. A number of techniques have been developed to measure the DNA content, based on sequential extraction blotting and hybridization with a labeled DNA probe. The sulphonate marker has been recently introduced by PBS-Orgenics; it allows the determination of picograms (pg) quantities of purified DNA. However, it is not simple to measure in complex biological samples especially when a large amount of protein is present. In considering the following points: 1. Precautions in handling the samples at different steps of preparation; 2. Modifications of the original technique 3. Concentration of samples expected at very low level, we are able to dose up to 2 pg of contaminant DNA per mg of MAb with a satisfactory reproducibility and reliability. This level is required not only to qualify final MAb but also to evaluate the efficiency of the purification process. Efforts are being made to achieve a better sensitivity.
authors.affiliation: Pasteur Mérieux Sérums et Vaccins, Immunology Research Department, Marcy L'Etoile, France.
authors.forename: J
authors.initials: J
authors.lastname: Carcagne
authors.forename: K T
authors.initials: KT
authors.lastname: Ha
authors.forename: M
authors.initials: M
authors.lastname: Latour
authors.forename: E D
authors.initials: ED
authors.lastname: Carosella
mesh_terms: D000911:Antibodies, Monoclonal / Q000302:isolation & purification* / Q000592:standards / Q000627:therapeutic use; D002478:Cells, Cultured; D003596:Cytosine / Q000031:analogs & derivatives; D004247:DNA / Q000032:analysis* / Q000187:drug effects; D004340:Drug Contamination; D006801:Humans; D011786:Quality Control; D013447:Sulfites / Q000494:pharmacology; D013451:Sulfonic Acids
chemical_list: D000911:Antibodies, Monoclonal; D013447:Sulfites; D013451:Sulfonic Acids; D003596:Cytosine; D004247:DNA; C009279:sodium bisulfite