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.

本研究的目的是确定消光系数 (EC) 的理论值与通过 Edelhoch 方法测量的大量生物治疗蛋白质的实验确定的消光系数相比如何。我们基于超过 176 项观察进行了广泛的分析，这些观察涵盖来自不同结构类别的 19 种不同类型的分子，包括 mAb、双特异性抗体、融合蛋白和 BiTE 分子。通过评估每个分子测量的可重复性并确定相对标准偏差 (%RSD) 来测量精度。对于任何给定分子，观察到的最大 RSD 为 1.7%，平均 RSD 为 0.9%。首先通过计算实验偏差来确定与理论消光系数的偏差，这是平均实验消光系数与理论消光系数之间的差值。然后将百分比偏差 (%bias) 计算为 (偏差 ÷ 理论 EC) × 100。观察到的任何给定分子的最大偏差百分比为 5.3%，平均偏差百分比为 2.6%。我们的结果表明，与氨基酸分析 (AAA) 技术等常用方法相比，Edelhoch 方法高度可靠，执行效率显着提高，成本、时间减少，安全性提高。