Case Study: The relief through customized manifolds
Avoiding product loss
Robustness / protection of single use bag
With RoSS® (Robust Storage and Shipping) the single use bag is covered by soft 3D foam, which provides a protective embedding of the single use bag and absorbs the expanding energy of the bag/liquid of about 8% due to the density drop during freezing. The heavy-duty plastic and stainless-steel lids withstand even the lowest temperatures and makes up the robust frame of RoSS®.
The implementation of RoSS® with the effect of a much more robust secondary packaging has diminished the risk of leakages or ruptures towards 0%. The single-use bag and its tubing are perfectly adapted in the RoSS® shell, which in turn maximizes the protective effect. After experiencing the freezing and shipment process with RoSS® for more than 500 times, the product loss has decreased from up to 5% down towards 0%.
“Full immobilization of the frozen single use bag guarantees that the bag, the tubing and all the accessories on the bag do not move at all. So, no breakage happens.”Johannes Kirchmair
Protein activity / potency
As protein activity should be granted even after scaling up the volumes, the target is to reach the same protein activity for different scales after freezing and/or thawing. A general approach for any protein or when comparing different proteins is not possible due to complex and protein-specific denaturation modes.
Sensitivity to denaturation at the ice liquid interface depends on the protein’s reaction. In collaborations, the DoE (Design of Experiment) regarding the effect of different controlled freezing rates and bag fill volumes was successfully tested on Laccase enzyme activity. Results showed that the controlled freezing in the example of plate freezers has a lower impact on the enzyme activity.1
With their slow and not very homogenous freezing processes, traditional static freezers lead to large crystalline structures that are the cause of friction, which in turn leads to the destruction of valuable proteins. Up to 20% of proteins can be destroyed like this. In some circumstances the loss of viral potency can rise to even 56%.2 Effect: After thawing, the substance has lost a considerable amount of its original concentration, thus dropping in value.3
Fast controlled freezing with plate-based freezer lead to a more homogeneous freezing with higher protein activity
Slow uncontrolled freezing (e.g. in static or blast freezer) decreases protein activity by occurring cryoconcentration.
Click here to download RoSS® brochure, case study product loss and sterility.
1 TU Wien: Freezing Analyse DoE im LabScale – Final Results, 2019. Data on file.
2 Bezavada A, et al.: Use of Blast Freezers in Vaccine Manufacturing. 2011. In: Bioprocess International https://bioprocessintl.com/manufacturing/monoclonal-antibodies/use-of-blast-freezers-in-vaccine-manufacture-322317/ [Accessed November 20, 2020]
3 Single Use Support: Challenges in Bulk Drug Substance Logistics: eBook. 2020. Available at: https://www.susupport.com/en/challenges-bulk-drug-substance-logistics/ [Accessed December 7, 2020]