Attribute Sciences, Amgen Inc., One Amgen Center Dr, M/S 30-1-B, Thousand Oaks, California, 91320, USA.
Coriolis Pharma, Martinsried, Munich, Germany.
Pharm Res. 2017 Dec;34(12):2817-2828. doi: 10.1007/s11095-017-2262-8. Epub 2017 Nov 6.
To physicochemically characterize and compare monoclonal antibody (mAb) solutions containing aggregates generated via metal catalyzed oxidation (MCO).
Two monoclonal IgG2s (mAb1 and mAb2) and one monoclonal IgG1 (rituximab) were exposed to MCO with the copper/ascorbic acid oxidative system, by using several different methods. The products obtained were characterized by complementary techniques for aggregate and particle analysis (from oligomers to micron sized species), and mass spectrometry methods to determine the residual copper content and chemical modifications of the proteins.
The particle size distribution and the morphology of the protein aggregates generated were similar for all mAbs, independent of the MCO method used. There were differences in both residual copper content and in chemical modification of specific residues, which appear to be dependent on both the protein sequence and the protocol used. All products showed a significant increase in the levels of oxidized His, Trp, and Met residues, with differences in extent of modification and specific amino acid residues modified.
The extent of total oxidation and the amino acid residues with the greatest oxidation rate depend on a combination of the MCO method used and the protein sequence.
对含有金属催化氧化（MCO）产生的聚集物的单克隆抗体（mAb）溶液进行物理化学表征和比较。
用铜/抗坏血酸氧化系统，通过几种不同的方法使两种单克隆 IgG2（mAb1 和 mAb2）和一种单克隆 IgG1（利妥昔单抗）暴露于 MCO 中。通过互补的聚集物和颗粒分析技术（从低聚物到微米级物质）以及质谱方法来确定残留铜含量和蛋白质的化学修饰，对获得的产物进行了表征。
所有 mAb 产生的蛋白质聚集物的粒径分布和形态相似，与所使用的 MCO 方法无关。残留铜含量和特定残基的化学修饰存在差异，这似乎取决于蛋白质序列和所用的方案。所有产物的氧化 His、Trp 和 Met 残基水平均显著升高，修饰程度和修饰的特定氨基酸残基存在差异。
总氧化程度和氧化速率最快的氨基酸残基取决于所使用的 MCO 方法和蛋白质序列的组合。
