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  • Journal article

  • Identification of novel glycosylation events on human serum-derived Factor IX

    • Authors : Pegg, Cassandra L; Zacchi, Lucia F; Recinos, Dinora Roche (2020)

  • Human Factor IX is a highly post-translationally modified protein that is an important clotting factor in the blood coagulation cascade. Functional deficiencies in Factor IX result in the bleeding disorder haemophilia B, which is treated with plasma-derived or recombinant Factor IX concentrates. Here, we investigated the post-translational modifications of human serum-derived Factor IX and report previously undescribed O-linked monosaccharide compositions at serine 141 and&...
  • Periodicals (Báo – Tạp chí)

  • Analysis of coagulation factor IX in bioreactor cell culture medium predicts yield and quality of the purified product

    • Authors : Zacchi, Lucia F; Recinos, Dinora Roche; Pegg, Cassandra L (2020)

  • Coagulation factor IX (FIX) is a highly complex post-translationally modified human serum glycoprotein and a high-value biopharmaceutical. The quality of recombinant FIX (rFIX), especially complete γ-carboxylation, is critical for rFIX clinical efficacy. Changes in bioreactor operating conditions can impact rFIX production and occupancy and structure of rFIX post-translational modifications (PTMs). We hypothesized that monitoring the bioreactor cell culture supernatant with Data Inde...
  • Journal article

  • S-Trap eliminates cell culture media polymeric surfactants for effective proteomic analysis of mammalian cell bioreactor supernatants

    • Authors : Zacchi, Lucia F; Recinos, Dinora Roche; Otte, Ellen (2020)

  • Proteomic analysis of bioreactor supernatants can inform on cellular metabolic status, viability, and productivity, as well as product quality, which can in turn help optimize bioreactor operation. Incubating mammalian cells in bioreactors requires the addition of polymeric surfactants such as Pluronic F68, which reduce the sheer stress caused by agitation. However, these surfactants are incompatible with mass spectrometry proteomics and must be eliminated during sampl...