Research and Early Development, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, United States.
Research and Early Development, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, United States.
J Pharm Biomed Anal. 2019 Feb 5;164:452-459. doi: 10.1016/j.jpba.2018.11.009. Epub 2018 Nov 5.
Maleimide functionalized polyethylene glycol (PEG) with varying degrees of branched chains is gaining increasing interest in drug conjugation. Characterization of these molecules in solution is highly challenging due to the multiplicity of the hydrolysis products and the subtle differences between them, especially for high molecular weight PEG-maleimide. A highly selective, sensitive and stability indicating HPLC method employing charged surface hybrid (CSH) stationary phase was developed for 8-arm PEG-maleimide characterization. The method has been validated per ICH Q2 guideline for specificity, sensitivity, linearity, accuracy and precision for quality and process control. Hydrolysis of 8-arm PEG-maleimide in different buffer media, pH and temperature was studied. The kinetics revealed the hydrolysis is a first-order reaction. Hydrolysis rate constant and half-life were obtained to guide protein conjugation process development.
支化程度不同的马来酰亚胺功能化聚乙二醇（PEG）在药物偶联中越来越受到关注。由于水解产物的多样性和它们之间的细微差异，特别是对于高分子量的 PEG-马来酰亚胺，对这些分子在溶液中的特性进行表征极具挑战性。本研究开发了一种高度选择性、灵敏且具有稳定性指示的 HPLC 方法，采用带电表面杂化（CSH）固定相来对 8 臂 PEG-马来酰亚胺进行表征。该方法已经按照 ICH Q2 指南进行了专属性、灵敏度、线性、准确度和精密度验证，可用于质量和工艺控制。研究了 8 臂 PEG-马来酰亚胺在不同缓冲介质、pH 值和温度下的水解情况。动力学研究表明，水解是一级反应。获得了水解速率常数和半衰期，以指导蛋白质偶联过程的开发。
