Department of Pharmacy, Pharmaceutical Technology and Biopharmaceutics, University of Munich, Butenandtstr. 5, 81377 Munich, Germany.
Sandoz Biopharmaceutics, Biochemiestr. 10, 6336 Langkampfen, Austria.
Int J Pharm. 2022 Jun 10;621:121760. doi: 10.1016/j.ijpharm.2022.121760. Epub 2022 Apr 30.
Biopharmaceutical products are subject to in depth analysis to ensure and improve their safety and efficacy. As part of this effort the stability and aggregation mechanisms of the therapeutic protein is characterized over the whole life cycle. The stability and aggregation behavior of single charge variants present in biopharmaceuticals were hardly investigated. In this study we applied a previously established methodology to assess the charge variants of the drug substance (DS) of human growth hormone (hGH). We assessed the stability and aggregation propensity of an acidic variant which forms in DS at a larger extent during short time storage at elevated temperatures. We developed a semi-preparative method to separate and analyze the charge species. Thermal and colloidal stability of this variant was analyzed by light scattering methods and a stability testing in different buffer formulations. The acidic variant showed slightly attractive self-interaction at lower pH. Thermal stress did not result in increased aggregation propensity or decreased stability compared to the DS. Thus, the methodology enabled to assess the risk of a single protein variant within the DS of hGH. The approach can also be utilized for other protein drugs as previously shown for a monoclonal antibody.
生物制药产品需要进行深入分析，以确保和提高其安全性和疗效。作为这项工作的一部分，对治疗性蛋白质在整个生命周期中的稳定性和聚集机制进行了表征。生物制药中存在的单电荷变异体的稳定性和聚集行为几乎没有得到研究。在这项研究中，我们应用了先前建立的方法来评估人生长激素（hGH）药物物质（DS）的电荷变异体。我们评估了在高温短时间储存过程中在 DS 中形成的酸性变异体的稳定性和聚集倾向。我们开发了一种半制备方法来分离和分析电荷种类。通过光散射方法和在不同缓冲液配方中的稳定性测试来分析这种变体的热和胶体稳定性。与 DS 相比，酸性变异体在较低 pH 值下表现出轻微的吸引力自相互作用。与热应力相比，它没有导致聚集倾向增加或稳定性降低。因此，该方法能够评估 hGH DS 中单个蛋白质变异体的风险。如前所述，该方法也可用于其他蛋白质药物，如单克隆抗体。
