Strategies for Bulk Storage and Shipment of Proteins

Jameel, Feroz; Padala, Chakradhar; Randolph, Theodore W.

doi:10.1002/9780470595886.ch27

Cited by 7 publications

(8 citation statements)

References 29 publications

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“…However, the previous authors focused on a horizontal freezing geometry that would not prevent the cryoconcentration heterogeneity discussed above. Jameel et al, in their review of bulk storage and shipment strategies, have discussed unidirectional freezing from the perspective of achieving a linearly scalable system and also for reducing cryoconcentration by promoting the formation of dendrites. However, they do not report on the ability to suppress density gradient driven convective flow as our work demonstrates.…”

Section: Introductionmentioning

confidence: 99%

The importance of heat flow direction for reproducible and homogeneous freezing of bulk protein solutions

Rodrigues¹,

Balzan²,

Rosa³

et al. 2013

Biotechnology Progress

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Freezing is an important operation in biotherapeutics industry. However, water crystallization in solution, containing electrolytes, sugars and proteins, is difficult to control and usually leads to substantial spatial solute heterogeneity. Herein, we address the influence of the geometry of freezing direction (axial or radial) on the heterogeneity of the frozen matrix, in terms of local concentration of solutes and thermal history. Solutions of hemoglobin were frozen radially and axially using small-scale and pilot-scale freezing systems. Concentration of hemoglobin, sucrose and pH values were measured by ice-core sampling and temperature profiles were measured at several locations. The results showed that natural convection is the major source for the cryoconcentration heterogeneity of solutes over the geometry of the container. A significant improvement in this spatial heterogeneity was observed when the freezing geometry was nonconvective, i.e., the freezing front progression was unidirectional from bottom to top. Using this geometry, less than 10% variation in solutes concentration was obtained throughout the frozen solutions. This result was reproducible, even when the volume was increased by two orders of magnitude (from 30 mL to 3 L). The temperature profiles obtained for the nonconvective freezing geometry were predicted using a relatively simple computational fluid dynamics model. The reproducible solutes distribution, predictable temperature profiles, and scalability demonstrate that the bottom to top freezing geometry enables an extended control over the freezing process. This geometry has therefore shown the potential to contribute to a better understanding and control of the risks inherent to frozen storage.

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Section: Introductionmentioning

confidence: 99%

The importance of heat flow direction for reproducible and homogeneous freezing of bulk protein solutions

Rodrigues¹,

Balzan²,

Rosa³

et al. 2013

Biotechnology Progress

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“…Frozen storage increases protein solution stability, extends the drug product shelf life, offers flexibility during manufacturing, mitigates the risk of microbial growth, and eliminates agitation and foaming during transportation (1)(2)(3). Thus, freezing and subsequent thawing (F/T) are integral steps in the manufacturing of most biopharmaceutical products (4).…”

Section: Introductionmentioning

confidence: 99%

“…It is important to ensure that the protein retains its biological activity and native state throughout manufacturing, distribution, and shelf life of the drug product. In this sense, non-native protein aggregates in the drug substance storage phase should be well controlled, as even small levels of aggregates can serve as nuclei for the formation of visible and subvisible particles when the solution is further exposed to fill and finish processing stresses (4).…”

Section: Introductionmentioning

confidence: 99%

“…CryoVessels® utilize stainless-steel vessels to F/T large volumes of drug substance (125 to 300 L). However, these systems are capital intensive, and large walk-in freezer rooms are required to store the frozen vessels (4,7). Additional operational challenges may arise after repeated F/T cycles due to surface modification of the stainless-steel vessels and increased levels of iron and other metals leaching into the protein solution (7).…”

Section: Introductionmentioning

confidence: 99%

“…An alternative type of controlled F/T system is the Celsius® CFT, which utilizes ethylene vinyl acetate (EVA) Celsius® bags containing up to 16.6 L of solution. These bags are compressed between two heat exchange plates through which a silicone-based heat transfer fluid is circulated (4). The frozen EVA film has a brittleness temperature in the range of −70 to −76°C; thus, there is a concern regarding its fragility during shipment and storage.…”

Section: Introductionmentioning

confidence: 99%

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Use of a Design of Experiments (DoE) Approach to Optimize Large-Scale Freeze-Thaw Process of Biologics

2021

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Large volumes of protein solutions are commonly stored in a frozen state before further drug product fill and finish. This study aimed to establish a design space to perform large-scale freeze-thaw (F/T) processes of biotherapeutics without inducing protein destabilization. A response surface model was designed to evaluate the following main factors and interactions: fill volume of the protein solution in 1-L containers, distance among nine containers during both F/T, freezer set temperature, and a novel forced air flow methodology during thawing. The analysis from 46 experimental runs indicated over 4-fold increase in the freezing rate by lowering the freezing temperature from −20 to −80°C, and the forced air flow at 98 fpm doubled the thawing rate. Furthermore, multivariate linear regression modeling revealed the significant impact of all main factors investigated on lactate dehydrogenase (LDH) quality attributes. The factor that most strongly affected the retention of LDH activity was the loading distance: ≥ 5 cm among containers positively affected the LDH activity response in 50.6%. The factor that most strongly retained the LDH tetramers was the set freezer temperature towards the lower range of −80°C (2.2% higher tetramer retention compared to −20°C freezing, due to faster freezing rate). In summary, this DoEbased systematic analysis increased F/T process understanding at large scale, identified critical F/T process parameters, and confirmed the feasibility of applying faster freezing and forced air thawing procedures to maintain the stability of LDH solutions subject to largescale F/T.

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Protein and solute distribution in drug substance containers during frozen storage and post‐thawing: A tool to understand and define freezing–thawing parameters in biotechnology process development

Kolhe

Badkar

2011

Biotechnology Progress

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Active pharmaceutical ingredient for biotechnology-based drugs, commonly known as drug substance (DS), is often stored frozen for longer shelf-life. Freezing DS enhances stability by slowing down reaction rates that lead to protein instability, minimizes the risk of microbial growth, and eliminates the risk of transport-related stress. High density polyethylene bottles are commonly used for storing monoclonal antibody DS due to good mechanical stress/strain resistant properties even at low temperatures. Despite the aforementioned advantages for frozen storage of DS, this is not devoid of risks. Proteins are known to undergo ice-water surface denaturation, cryoconcentration, and cold denaturation during freezing. A systematic investigation was performed to better understand the protein and solute distribution along with potential of aggregate formation during freeze and thaw process. A significant solute and protein concentration gradient was observed for both frozen and thawed DS bottles. In case of thawed DS, cryoconcentration was localized in the bottom layer and a linear increase in concentration as a function of liquid depth was observed. On the other hand, for frozen DS, a "bell shaped" cryoconcentration distribution was observed between the bottom layers and centre position. A cryoconcentration of almost three-fold was observed for frozen DS in the most concentrated part when freezing was conducted at -20 and -40 °C and 2.5-fold cryoconcentration was observed in the thawed DS before mixing. The information obtained in this study is critical to design freeze thaw experiments, storage condition determination, and process improvement in manufacturing environment.

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Strategies for Bulk Storage and Shipment of Proteins

Cited by 7 publications

References 29 publications

The importance of heat flow direction for reproducible and homogeneous freezing of bulk protein solutions

The importance of heat flow direction for reproducible and homogeneous freezing of bulk protein solutions

Use of a Design of Experiments (DoE) Approach to Optimize Large-Scale Freeze-Thaw Process of Biologics

Protein and solute distribution in drug substance containers during frozen storage and post‐thawing: A tool to understand and define freezing–thawing parameters in biotechnology process development

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