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Investigations into, and development of, a lyophilized and formulated recombinant human factor IX produced from CHO cells

Almeida, Aline G., Pinto, Rodrigo C. V., Smales, Christopher Mark, Castilho, Leda R. (2017) Investigations into, and development of, a lyophilized and formulated recombinant human factor IX produced from CHO cells. Biotechnology Letters, 39 (8). pp. 1109-1120. ISSN 0141-5492. E-ISSN 1573-6776. (doi:10.1007/s10529-017-2353-y) (KAR id:61688)

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Objectives: To develop a recombinant human factor IX (rFIX) formulation equivalent to commercially available products in terms of cake appearance, residual moisture, proportion of soluble aggregates and activity maintenance for 3 months at 4–8 °C.

Results: NaCl and low bulking agent/cryoprotectant mass ratio had a negative impact on cake quality upon lyophilisation for a wide range of formulations tested. Particular devised formulations maintained rFIX activity after lyophilization with a similar performance when compared with the rFIX formulated using the excipients reported for a commercially available FIX formulation (Benefix). rFIX remained active after 3 months when stored at 4 °C, though this was not the case with samples stored at 40 °C. Interestingly, particular formulations had an increase in residual moisture after 3 months storage, but not above a 3% threshold. All four formulations tested were equivalent to the Benefix formulation in terms of particle size distribution and cake appearance.

Conclusions: Three specific formulations, consisting of surfactant polysorbate-80, sucrose or trehalose as cryoprotectant, mannitol or glycine as bulking agent, l-histidine as buffering agent, and NaCl added in the reconstitution liquid at 0.234% (w/v) were suitable for use with a CHO cell-derived recombinant FIX.

Item Type: Article
DOI/Identification number: 10.1007/s10529-017-2353-y
Uncontrolled keywords: Excipients Formulation Freeze-drying Lyophilization Recombinant factor IX Stability study
Divisions: Divisions > Division of Natural Sciences > Biosciences
Depositing User: Mark Smales
Date Deposited: 11 May 2017 08:21 UTC
Last Modified: 29 May 2019 19:02 UTC
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