Sterilization of a blood derived products using supercritical carbon dioxide (SCCO2)

Research output: Types of ThesisMaster's Thesis


Human platelet lysate (hPL), has a myriad of molecules like growth factors, and it is used mainly as a cell culture growth supplement. The increase in interest in the product could be related to several European legislation that strengthened the importance of the use of non-animal methods. As a result, standardization of human platelet lysate production is a requirement to achieve batch to batch consistency for commercialization. From the several parameters considered inactivation/removal of viruses methods are essential. Thus, sterilization technologies, processes that focus on the complete elimination, removal or inactivation of all forms of microbial life, can be implemented to comply with high standards of safety. Current standard technologies such as moist and dry heat sterilization, ethylene oxide and gamma radiation are typically deleterious to biological material. Recently, supercritical carbon dioxide (CO2) presents as a solution for the sterilization of biological products. As such, the present thesis focused on demonstrating the effectiveness of supercritical CO2 technology as a viable sterilization alternative for lyophilized human platelet lysate and studying the chemical and biochemical properties of the final product to evaluate the impact on its composition. For this purpose, the experimental work that was divided into the pilot and main study. The supercritical carbon dioxide protocol (4h cycle, 40ºC, 140bar and 300 ppm of hydrogen peroxide) applied to human platelet lysate was implemented and analyzed. Parameters for the experimental work were adjusted between the pilot and main study. Results revealed that the protocol proposed was not able to sterilize the hPL samples. The supercritical CO2 protocol applied induced a loss of total protein content, epidermal growth factor (EGF) and platelet-derived growth factor-BB (PDGF-BB), less than 14%, approximately 11%, and about 17%, respectively, when compared with untreated samples. The small content decrease is inferior to the loss reported by others work that used different inactivation or sterilization protocols. The pH value was also not affected by the sterilization protocol. Since the protocol applied was not optimized for the packaging used analysis on the topic was performed. The results showed that indeed the packaging could interfere with the process. Consequently, the technology applied presents the potential for effective sterilization of hPL opening possibility for future protocol optimization to achieve terminal sterilization.
Original languageEnglish
  • Oliveira, Ana Leite, Supervisor
  • Ribeiro, Nilza, Supervisor
Award date7 Nov 2019
Publication statusPublished - 7 Nov 2019


  • Human platelet lysate
  • Growth factors
  • Total protein
  • Epidermal growth factor
  • Platelet-derived growth factor-BB
  • Supercritical CO2 technolog


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