Amgen, Inc. Thousand Oaks, CA, USA.
Amgen, Inc. Thousand Oaks, CA, USA.
Biologicals. 2021 Jun;71:42-47. doi: 10.1016/j.biologicals.2021.03.003. Epub 2021 Apr 17.
The objective of this study was to determine how the theoretical values of the extinction coefficient (EC) compares to the experimentally determined extinction coefficient for a large set of biotherapeutic proteins measured by the Edelhoch method. We have performed extensive analysis based on over 176 observations covering 19 different types of molecules from different structural classes covering mAbs, bispecific antibodies, fusion proteins and BiTE molecules. Precision was measured by assessing the repeatability of the measurements for each molecule and determining the relative standard deviation (%RSD). The maximum RSD observed for any given molecule was 1.7% with an average RSD of 0.9%. Deviation from the theoretical extinction coefficient was determined by calculating the experimental bias first, which is the difference between the mean experimental extinction coefficient and the theoretical extinction coefficient. The percent bias (%bias) was then calculated as (bias ÷ theoretical EC) × 100. The maximum %bias observed for any given molecule was 5.3% with an average %bias of 2.6%. Our results indicate that the Edelhoch method is highly reliable with significant improvement in execution efficiency with reduction in cost, time and improvements in safety when compared to the commonly used methods such as amino acid analysis (AAA) technique.
本研究旨在确定理论消光系数 (EC) 值与通过 Edelhoch 法测量的大量生物治疗蛋白的实验确定消光系数之间的关系。我们基于超过 176 个观察结果进行了广泛的分析，涵盖了来自不同结构类别（包括单克隆抗体、双特异性抗体、融合蛋白和 BiTE 分子）的 19 种不同类型的分子。通过评估每个分子的测量重复性并确定相对标准偏差 (%RSD) 来衡量精密度。任何给定分子的最大 RSD 为 1.7%，平均 RSD 为 0.9%。通过首先计算实验偏差来确定与理论消光系数的偏差，实验偏差是平均实验消光系数与理论消光系数之间的差异。然后将偏差百分比 (%bias) 计算为 (偏差 ÷ 理论 EC) × 100。任何给定分子的最大 %bias 为 5.3%，平均 %bias 为 2.6%。我们的结果表明，与常用方法（如氨基酸分析 (AAA) 技术）相比，Edelhoch 法具有高度可靠性，在执行效率方面有显著提高，同时降低了成本、时间并提高了安全性。
