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| SNS:An Effective Strategy for Extending Ceramic Hydroxyapatite Column Life |
| DENG Yi-xi1, LI Ji-dong2, LI Le1, MENG Guo-ji1, YU Yu-gen1 |
1. Shenzhen Main Luck Pharmaceuticals Inc., Shenzhen 518029, China;
2. Bio-Rad Laboratories(Shanghai) Ltd., Shenzhen 510095, China |
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Abstract Ceramic Hydroxyapatite (CHT) has been widely used in the manufacturing of a wide variety of monoclonal antibodies (mAbs), vaccines, and other protein therapeutics. It is an effective tool for the removal of residual host cell proteins, nucleic acids, viruses, endotoxins, and product-related contaminants such as aggregates and degradation fragments. However, damage of the CHT media and increased column back pressure are often observed at production scale-up. A transient pH excursion during elution with increased salt concentration leads to Ca2+ depletion from the CHT column. In the present study,the effect of surface neutralization system (SNS) on the life cycles of CHT media is demonstrated. Following post-loading wash, a buffer with slightly elevated pH is applied onto the column so as to prevent significant pH drop during target protein elution. The level of Ca2+ depletion in response to pH and salt concentration of the SNS buffer have been investigated, and the optimal buffer composition for maintaining the pH of column effluent near neutral been identified. This newly developed protocol has been validated for the polish purification of mAb WLB304. The dynamic binding capacity of WLB304, impurity clearance efficiency, leached Ca2+ level, pH excursion and column back pressure during purification process were closely monitored. Data indicate that the implementation of SNS can significantly reduce Ca2+ depletion and effectively extend the life time of a CHT column, without compromising its chromatography performance.
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Received: 25 July 2016
Published: 25 January 2017
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