一种新的评价技术，用于通过 CE 与 native UV-LIF 检测纯化蛋白质中的杂质。

A new evaluation technique for the detection of impurities in purified proteins via CE with native UV-LIF.

机构信息

Laboratoire des IMRCP, Université de Toulouse, Université Paul Sabatier, Toulouse, France.

出版信息

Electrophoresis. 2010 Jan;31(2):396-402. doi: 10.1002/elps.200900369.

DOI:10.1002/elps.200900369

PMID:20084633

Abstract

An analytical methodology for quality control analyses of IgG and their impurities is presented using a new UV-LIF (266 nm) detector inside the cassette of a CE instrument and its performance was evaluated. The observed sensitivity was very close to that obtained by silver staining of slab gels (LOD of 25 ng/mL), while the sensitivity of the analysis is 80 times better than with CE/UV absorption (214 nm). Examples of the analysis of pharmaceutical and other commercial IgGs are provided and the kinetics of the reduction of IgG by beta-mercaptoethanol is reported, demonstrating the ease of performing the analysis.

摘要

介绍了一种使用 CE 仪器盒内新型 UV-LIF（266nm）检测器进行 IgG 及其杂质质量控制分析的分析方法，并对其性能进行了评估。观察到的灵敏度非常接近平板凝胶银染（LOD 为 25ng/mL）获得的灵敏度，而分析灵敏度比 CE/UV 吸收（214nm）好 80 倍。提供了药物和其他商业 IgG 分析的实例，并报告了β-巯基乙醇还原 IgG 的动力学，证明了进行分析的容易性。