Lin Joyce, Lazar Alexandru C
Analytical & Pharmaceutical Sciences, ImmunoGen, Inc., Waltham, MA, USA.
Methods Mol Biol. 2013;1045:295-302. doi: 10.1007/978-1-62703-541-5_19.
Imaged capillary isoelectric focusing (icIEF) is capable of monitoring the charge heterogeneity profile of conjugated antibodies. The electropherogram from icIEF can be integrated to quantitate the amount of unconjugated antibody present in a conjugate sample. This chapter describes an icIEF method where a conjugate sample was first prepared by mixing with appropriate ampholytes, pI markers, and additives. Then, the sample was focused in a fluorocarbon-coated fused silica capillary, where absorbance images were taken. Quantitation of the unconjugated antibody was achieved by using a calibration curve.
成像毛细管等电聚焦（icIEF）能够监测偶联抗体的电荷异质性谱。icIEF的电泳图可用于定量偶联物样品中未偶联抗体的含量。本章介绍了一种icIEF方法，首先将偶联物样品与合适的两性电解质、等电点标记物和添加剂混合制备样品。然后，将样品在涂有氟碳的熔融石英毛细管中聚焦，并采集吸光度图像。通过使用校准曲线实现未偶联抗体的定量。
