Powers Thomas W, Mariani Shawn, Narepekha Halyna, Ryan Daniel, Sankar Savita, Lerch Thomas F
Pfizer Inc., Analytical Research and Development, 875 Chesterfield Pkwy. West, Chesterfield, MO 63017, USA.
Mol Ther Methods Clin Dev. 2025 Aug 13;33(3):101560. doi: 10.1016/j.omtm.2025.101560. eCollection 2025 Sep 11.
The multi-attribute method (MAM), a mass spectrometry technique for quantifying amino acid modifications at the peptide level, is becoming a prominent analytical tool in the development of biotherapeutics. The method has promise for adeno-associated virus (AAV) therapeutics, where capsid protein modifications have been directly linked to reduced transduction efficiency. Given this link, a robust and precise procedure to quantitate capsid modifications would be beneficial for implementation throughout biotherapeutic development. Herein, an AAV product was characterized, and capsid sequence liabilities were identified. A peptide map MAM method was developed to quantitate select sites of modifications and was validated according to ICH Q2(R2). Through this exercise, the method was demonstrated to be suitable to quantitate several sites of deamidation and the method was applied during stability, process development, and product comparability studies. Additionally, preliminary data demonstrated that the method was not limited to monitoring deamidation but also could be applied to other post-translational and chemical modifications.
多属性方法（MAM）是一种用于在肽水平定量氨基酸修饰的质谱技术，正成为生物治疗药物开发中一种突出的分析工具。该方法在腺相关病毒（AAV）治疗方面具有前景，在AAV治疗中，衣壳蛋白修饰已被直接证明与转导效率降低有关。鉴于这种联系，一种强大而精确的定量衣壳修饰的方法对于整个生物治疗药物开发过程的实施将是有益的。在此，对一种AAV产品进行了表征，并确定了衣壳序列缺陷。开发了一种肽图MAM方法来定量选定的修饰位点，并根据ICH Q2(R2)进行了验证。通过这项工作，证明该方法适用于定量多个脱酰胺位点，并且该方法已应用于稳定性、工艺开发和产品可比性研究。此外，初步数据表明该方法不仅限于监测脱酰胺，还可应用于其他翻译后修饰和化学修饰。
