Biotechnology Letters

, Volume 39, Issue 8, pp 1109–1120 | Cite as

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

  • Aline G. Almeida
  • Rodrigo C. V. Pinto
  • C. Mark Smales
  • Leda R. Castilho
Original Research Paper

Abstract

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.

Keywords

Excipients Formulation Freeze-drying Lyophilization Recombinant factor IX Stability study 

Supplementary material

10529_2017_2353_MOESM1_ESM.docx (742 kb)
Supplementary material 1 (DOCX 742 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  1. 1.Cell Culture Engineering Laboratory, COPPE, PEQFederal University of Rio de Janeiro (UFRJ)Rio de JaneiroBrazil
  2. 2.IQ, Biochemistry ProgramFederal University of Rio de Janeiro (UFRJ)Rio de JaneiroBrazil
  3. 3.FIOCRUZ, BiomanguinhosRio De JaneiroBrazil
  4. 4.Industrial Biotechnology Centre and School of BiosciencesUniversity of Kent at CanterburyCanterburyUK

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