Drug Product Science & Technology, Bristol-Myers Squibb, Co., 1 Squibb Drive, New Brunswick, New Jersey, 08901, USA.
AAPS PharmSciTech. 2017 Apr;18(3):803-808. doi: 10.1208/s12249-016-0570-7. Epub 2016 Jun 24.
Formation of isoaspartate (IsoAsp) from spontaneous asparagine (Asn) deamidation or aspartate (Asp) isomerization is one of the most common non-enzymatic pathways of chemical degradation of protein and peptide pharmaceuticals. Rapid quantitation of IsoAsp formation can enable rank-ordering of potential drug candidates, mutants, and formulations as well as support shelf life prediction and stability requirements. A coupled enzymatic fluorescence-based IsoAsp assay (CEFIA) was developed as a high-throughput method for quantitation of IsoAsp in peptides and proteins. In this note, application of this method to two therapeutic candidate proteins with distinct structural scaffolds is described. In addition, the results obtained with this method are compared to those from conventional assays.
异天冬氨酸（IsoAsp）的形成是蛋白质和肽类药物最常见的非酶化学降解途径之一，它来自自发天冬酰胺（Asn）脱酰胺或天冬氨酸（Asp）异构化。快速定量异天冬氨酸的形成可以对潜在候选药物、突变体和制剂进行排序，并支持保质期预测和稳定性要求。本文描述了一种基于酶联荧光的异天冬氨酸检测方法（CEFIA），该方法可用于定量检测肽和蛋白质中的异天冬氨酸。此外，还将该方法的结果与传统方法进行了比较。
