Impact of Buffer, Protein Concentration and Sucrose Addition on the Aggregation and Particle Formation during Freezing and Thawing

• Published in: Pharmaceutical research Vol. 35; no. 5; pp. 101 - 16
• Main Authors: Hauptmann, Astrid, Podgoršek, Katja, Kuzman, Drago, Srčič, Stanko, Hoelzl, Georg, Loerting, Thomas
• Format: Journal Article
• Language: English
• Published: New York Springer US 01-05-2018; Springer; Springer Nature B.V
• Subjects: Antibodies, Monoclonal - chemistry; Biochemistry; Biomedical and Life Sciences; Biomedical Engineering and Bioengineering; Biomedicine; Buffers; Calorimetry; Calorimetry, Differential Scanning; Chemistry, Pharmaceutical; Cryoprotectors; Crystallization; Differential scanning calorimetry; Drug Compounding - methods; Drug Stability; Drug Storage; Freeze-thawing; Freezing; Freezing - adverse effects; Health aspects; Immunologic Factors - chemistry; Light scattering; Medical Law; Monoclonal antibodies; Morphology; Pharmacology/Toxicology; Pharmacy; Photon correlation spectroscopy; Protein Aggregates; Protein Stability; Proteins; Research Paper; Sucrose; Sucrose - chemistry; Sugar; Temperature; Thawing; thawing; cryomicroscopy; freezing; buffer and mAb formulation; particle formation
• ISSN: 0724-8741; 1573-904X
• DOI: 10.1007/s11095-018-2378-5

Purpose
This study addresses the effect of freezing and thawing on a therapeutic monoclonal antibody (mAb) solution and the corresponding buffer formulation. Particle formation, crystallization behaviour, morphology changes and cryo-concentration effects were studied after varying the freezing and thawing rates, buffer formulation and protein concentration. The impact of undergoing multiple freeze/thaw (FT)-cycles at controlled and uncontrolled temperature rates on mAb solutions was investigated in terms of particle formation.
Methods
Physicochemical characteristics were analysed by Differential Scanning Calorimetry whereas morphology changes are visualized by cryomicroscopy measurements. Micro Flow Imaging, Archimedes and Dynamic Light Scattering were used to investigate particle formation.
Results
Data retrieved in the present study emphasizes the damage caused by multiple FT-cyles and the need for sucrose as a cryoprotectant preventing cold-crystallization specifically at high protein concentrations. Low protein concentrations cause an increase of micron particle formation. Low freezing rates lead to a decreased particle number with increased particle diameter.
Conclusion
The overall goal of this research is to gain a better understanding of the freezing and thawing behaviour of mAb solutions with the ultimate aim to optimize this process step by reducing the unwanted particle formation, which also includes protein aggregates.