Formulation and Analytical Resources, Amgen, Inc., One Amgen Center Drive, Thousand Oaks, California 91320, USA.
J Pharm Sci. 2009 Dec;98(12):4695-710. doi: 10.1002/jps.21778.
Tungsten has been associated with protein aggregation in prefilled syringes (PFSs). This study probed the relationship between PFSs, tungsten, visible particles, and protein aggregates. Experiments were carried out spiking solutions of two different model proteins with tungsten species obtained from the extraction of tungsten pins typically used in syringe manufacturing processes. These results were compared to those obtained with various soluble tungsten species from commercial sources. Although visible protein particles and aggregates were induced by tungsten from both sources, the extract from tungsten pins was more effective at inducing the formation of the soluble protein aggregates than the tungsten from other sources. Furthermore, our studies showed that the effect of tungsten on protein aggregation is dependent on the pH of the buffer used, the tungsten species, and the tungsten concentration present. The lower pH and increased tungsten concentration induced more protein aggregation. The protein molecules in the tungsten-induced aggregates had mostly nativelike structure, and aggregation was at least partly reversible. The aggregation was dependent on tungsten and protein concentration, and the ratio of these two and appears to arise through electrostatic interaction between protein and tungsten molecules. The level of tungsten required from the various sources was different, but in all cases it was at least an order of magnitude greater than the typical soluble tungsten levels measured in commercial PFS.
钨已被证实与预充式注射器（PFS）中的蛋白质聚集有关。本研究探究了 PFS、钨、可见颗粒和蛋白质聚集体之间的关系。实验将两种不同模型蛋白的溶液与从典型用于注射器制造过程的钨针提取的钨物种混合。将这些结果与从商业来源获得的各种可溶性钨物种的结果进行了比较。尽管来自两种来源的钨都能诱导可见的蛋白质颗粒和聚集体，但来自钨针的提取物比其他来源的钨更能有效地诱导可溶性蛋白质聚集体的形成。此外，我们的研究表明，钨对蛋白质聚集的影响取决于所用缓冲液的 pH 值、钨物种和存在的钨浓度。较低的 pH 值和增加的钨浓度诱导更多的蛋白质聚集。钨诱导的聚集体中的蛋白质分子大多具有天然结构，且聚集至少部分是可逆的。聚集取决于钨和蛋白质浓度，这两个浓度的比例似乎是通过蛋白质和钨分子之间的静电相互作用产生的。来自各种来源的钨所需的水平不同，但在所有情况下，它至少比商业 PFS 中测量的典型可溶性钨水平高一个数量级。
