caa a22 4500 477127851 CHVBK 20180405111655.0 cr unu---uuuuu 170330e19970801xx s 000 0 eng 10.1023/A:1012169832299 doi (NATIONALLICENCE)springer-10.1023/A:1012169832299 Evaluation of Genetic Stability of Recombinant Human Factor VIII by Peptide Mapping and On-line Mass Spectrometric Analysis [Elektronische Daten] [Marc Besman, Donald Shiba] Purpose. The genetic stability of a recombinant human factor VIII (rhFVIII) product expressed in Chinese hamster ovary cells (Recombinate™) has been evaluated through comparisons of the protein produced at the beginning, middle and end of a typical production campaign. Methods. Recombinant human factor VIII was incubated with thrombin, the resulting four polypeptides were isolated by RP-HPLC, subjected to proteolysis with trypsin, and the peptide mixtures were resolved by RP-HPLC. Tryptic peptide mixtures were subjected to online mass spectrometric analysis using an electrospray ionization source interfaced to a quadrupole mass analyzer scanning from 1950−200 amu, and the peptide ion data were compared for three lots produced from the beginning, middle and end of a production campaign. Results. The UV elution profiles for each of the rhFVIIIa polypeptides were highly similar for factor VIII isolated from the beginning, middle and end of production. Total ion data from the peptide maps derived from three lots of rhFVIII were compared by MH1+ values as a function of scan range. A total of 918 ions were analyzed for the four polypeptides of rhFVIII produced at the beginning, middle and end of a production campaign. The ions were detected at the same relative retention times, as indicated by the similar scan numbers for the three lots. Conclusions. These observations support that rhFVIII preparations produced from the beginning, middle and end of a production campaign were highly similar, and demonstrate genetic stability in the manufacturing process of Recombinate™. Plenum Publishing Corporation, 1997 factor VIII nationallicence genetic stability nationallicence peptide mapping nationallicence mass spectrometry nationallicence proteolysis nationallicence Besman Marc Hyland Division, Baxter Healthcare, Biotech Group, 1720 Flower Avenue, 91010, Duarte, California aut Shiba Donald Hyland Division, Baxter Healthcare, Biotech Group, 1720 Flower Avenue, 91010, Duarte, California aut Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/8(1997-08-01), 1092-1098 0724-8741 14:8<1092 1997 14 11095 https://doi.org/10.1023/A:1012169832299 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1012169832299 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Besman Marc Hyland Division, Baxter Healthcare, Biotech Group, 1720 Flower Avenue, 91010, Duarte, California aut NATIONALLICENCE 700 1- Shiba Donald Hyland Division, Baxter Healthcare, Biotech Group, 1720 Flower Avenue, 91010, Duarte, California aut NATIONALLICENCE 773 0- Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/8(1997-08-01), 1092-1098 0724-8741 14:8<1092 1997 14 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer