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Monitoring ceramic hydroxyapatite media degradation using dynamic image analysis and uniaxial confined bulk
Oliver Kaltenbrunner1, Shawn Cao, Esteban Freydell
1Purification Process Development, Amgen Inc., Thousand Oaks, CA 91320, USA. oliverk@amgen.com
Biotechnology Journal
|June 8, 2012
Summary
Adding phosphate and/or calcium to buffers minimizes degradation of ceramic hydroxyapatite (CHT) chromatography media under acidic conditions. These additives preserve CHT particle solidity and mechanical strength, enhancing packed bed lifetime.
Area of Science:
- Biochemistry
- Materials Science
- Chemical Engineering
Background:
- Ceramic hydroxyapatite (CHT) is a key chromatographic medium for biomolecule purification in the pharmaceutical industry.
- CHT exhibits moderate stability under acidic conditions, potentially limiting packed bed lifetime and performance.
- Monitoring CHT degradation is crucial for optimizing buffer systems and ensuring process robustness.
Purpose of the Study:
- To investigate the application of dynamic image analysis (DIA) and uniaxial confined bulk compression (UCBC) for monitoring CHT particle degradation.
- To evaluate the impact of buffer composition on CHT particle properties, including solidity, morphology, and mechanical strength.
- To determine optimal buffer conditions for preserving CHT integrity during chromatography.
Main Methods:
- Dynamic Image Analysis (DIA) was employed to assess changes in particle solidity and morphology.
- Uniaxial Confined Bulk Compression (UCBC) was utilized to measure alterations in resistance to compression.
- CHT particle properties were analyzed as a function of buffer composition, bed position, and operational parameters.
Main Results:
- Exposure to acidic pH induced degradation in CHT particles, affecting solidity and mechanical strength.
- The addition of 1 mM phosphate and/or calcium to acidic buffers significantly minimized CHT particle degradation.
- Calcium and phosphate ions positively influenced CHT mechanical behavior and particle solidity, preserving morphological characteristics.
Conclusions:
- Phosphate and calcium supplementation in buffers effectively mitigates CHT degradation under acidic chromatographic conditions.
- DIA and UCBC are valuable diagnostic tools for assessing CHT stability and guiding the design of robust purification processes.
- Optimized buffer systems incorporating calcium and phosphate can enhance the longevity and performance of CHT packed beds in biomolecule purification.
