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摘要
A commercial kit, consisting of magnetic particle-based DNA extraction, followed by a quantitative PCR analysis, was commonly used to detect the residual host DNA of biopharmaceuticals made in CHO cells. However, the recovery of the target DNA was just no more than 30% which is much lower than that the regulatory guideline expected between 80% to 120% range, especially when the kit was applied to the residual CHO DNA detection in monoclonal antibody samples. In this study, we describe our improvements in the recovery on extracting residual DNA from monoclonal antibody samples that often contains high protein concentration.
Enzyme/protein ratio, incubate conditions and elution conditions were developed and the optimized method resulted in the increasing of DNA recovery from 30% to 80% and above. Besides, high speed centrifugation turned out to help lowering the particle loss risk and shortening the time of the magnetic particles attracted by the magnetic stand. The optimized method was validated formally to identify its accuracy, precision and robustness. At last, the validated method was applied to samples analysis from different purification processes as well as the final drug product release successfully.
- 出版日期2015-6
- 单位上海医药工业研究院
