Department of Bioanalytical Sciences, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
Bioanalysis. 2020 Dec;12(23):1671-1679. doi: 10.4155/bio-2020-0191. Epub 2020 Nov 12.
PEGylation technology is one of long-acting delivery (LAD) platforms used to increase half-life of protein therapeutics. However, PEGylation of anti-Factor D Fab (PEG-aFD) poses challenges for detecting anti-drug antibody (ADA) to both Fab and polyethylene glycol (PEG) portions. Although the bridging ELISA using traditional assay diluent containing Tween 20 is good for detecting ADA to Fab, it failed to detect ADA to PEG. Instead of only reducing Tween 20 in assay diluent, using a proprietary commercial buffer PEG50-1 as assay diluent successfully enabled the detection of ADA to both Fab and PEG with fit-for-purpose sensitivity and drug tolerance. Identification of appropriate assay diluent is critical for detection of ADA to both Fab and PEG in a PEGylated molecule.
聚乙二醇化技术是一种长效递药（LAD）平台，用于延长蛋白类治疗药物的半衰期。然而，抗因子 D Fab 的聚乙二醇化（PEG-aFD）对 Fab 和聚乙二醇（PEG）部分的抗药物抗体（ADA）检测构成了挑战。尽管使用含有吐温 20 的传统检测稀释液的桥联 ELISA 非常适合检测针对 Fab 的 ADA，但它未能检测针对 PEG 的 ADA。替代方法是，在检测稀释液中不仅减少吐温 20 的含量，而且使用专有的商业缓冲液 PEG50-1 作为检测稀释液，成功地实现了针对 Fab 和 PEG 的 ADA 的检测，具有适合目的的灵敏度和药物耐受性。对于 PEG 化分子中针对 Fab 和 PEG 的 ADA 的检测，鉴定合适的检测稀释液至关重要。
