Characterization of a Laboratory-Scale Container for Freezing Protein Solutions with Detailed Evaluation of a Freezing Process Simulation
Characterization of a Laboratory-Scale Container for Freezing Protein Solutions with Detailed Evaluation of a Freezing Process Simulation
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DOI：
10.1002/jps.23814
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发表时间：
2014-02-01
影响因子：
3.8
通讯作者：
Nidetzky, Bernd
中科院分区：
文献类型：
--
作者：
Roessl, Ulrich;Jajcevic, Dalibor;Nidetzky, Bernd
构建了一个300 mL不锈钢冷冻容器，以实现符合QbD（设计质量）的研究和优化中等体积蛋白质药物的冷冻和解冻（F/T）过程。冷冻性能的表征进行冷冻动力学，温度曲线，冷冻浓缩，和稳定性的冷冻蛋白质。建立了温度和相变的计算流体动力学（CFD）模拟，以促进过程缩放和过程分析以及未来冷冻容器的定制。使用牛血清白蛋白从冰芯样品中测定蛋白质冷冻浓缩。研究了乳酸脱氢酶（LDH）在冻存兔肌中的活性、聚集和结构扰动。计算流体力学模拟提供了良好的定性和定量的协议与高分辨率的温度和相变的实验测量，也允许空间cryoconcentration模式的估计。LDH在实验室规模的系统中表现出对冷冻的稳定性，表明在一定条件下冷冻浓缩的保护作用。实验室规模的冷冻容器与精确的CFD建模相结合，将允许在先进的规模下对F/T过程进行更深入的研究，从而代表了更好地理解过程的重要一步。(C)2013 Wiley Periodicals，Inc.
A 300-mL stainless steel freeze container was constructed to enable QbD (Quality by Design)-compliant investigations and the optimization of freezing and thawing (F/T) processes of protein pharmaceuticals at moderate volumes. A characterization of the freezing performance was conducted with respect to freezing kinetics, temperature profiling, cryoconcentration, and stability of the frozen protein. Computational fluid dynamic (CFD) simulations of temperature and phase transition were established to facilitate process scaling and process analytics as well as customization of future freeze containers. Protein cryoconcentration was determined from ice-core samples using bovine serum albumin. Activity, aggregation, and structural perturbation were studied in frozen rabbit muscle L-lactic dehydrogenase (LDH) solution. CFD simulations provided good qualitative and quantitative agreement with highly resolved experimental measurements of temperature and phase transition, allowing also the estimation of spatial cryoconcentration patterns. LDH exhibited stability against freezing in the laboratory-scale system, suggesting a protective effect of cryoconcentration at certain conditions. The combination of the laboratory-scale freeze container with accurate CFD modeling will allow deeper investigations of F/T processes at advanced scale and thus represents an important step towards a better process understanding. (C) 2013 Wiley Periodicals, Inc.