Abstract：
An imaged capillary isoelectric focusing (icIEF)-based method was developed and validated as a multi-attribute method for a bispecific antibody (BsAb). First, as the traditional application of the icIEF method, it serves as an identity assay and purity assay for the BsAb. Second, the method can also be used as an impurity assay for the homodimer monoclonal antibodies generated during BsAb assembly. The homodimer impurity analysis for BsAb is usually done by hydrophobic interaction chromatography methods in the industry. The icIEF method has good sensitivity (down to 4 µg/mL in a limit of quantitation) when UV fluorescence detection is used, which detects the native fluorescence of proteins. This is the first report that an icIEF method has been applied as impurity assay.
摘要：
开发了一种基于成像毛细管等电聚焦（icIEF）的方法，并将其验证为双特异性抗体（BsAb）的多属性方法。首先,作为ICIEF方法的传统应用，它用作BsAb的身份测定和纯度测定。第二,该方法也可用作BsAb组装过程中产生的同二聚体单克隆抗体的杂质测定。BsAb的同二聚体杂质分析通常通过工业中的疏水相互作用色谱法进行。当使用UV荧光检测时，icIEF方法具有良好的灵敏度（定量极限低至4µg/mL），检测蛋白质的天然荧光。这是首次报道icIEF方法已用作杂质测定。
