Zhou Chao-Ming
Department of Quality Research, Shanghai Zerun Biotechnology Co., Ltd, Shanghai, China.
Electrophoresis. 2013 Nov;34(20-21):3046-53. doi: 10.1002/elps.201300161. Epub 2013 Oct 1.
CIEF with whole-column imaging detection (WCID) can be a useful tool for the characterization and identification of human papillomavirus (HPV). This article is the initial report of the determination of the pI of HPV by CIEF-WCID method. In this study, components of the assay selected for optimization were ampholytes, additives, methylcellulose concentration, HPV concentration, salt concentration, and focusing time. Then the optimization CIEF-WCID method was validated for HPV 16L1 and HPV 18L1. As a result, a precise method to analyze the pI values of HPVs was achieved with RSD < 1.0%. The HPV peak pattern was reproducible. CIEF-WCID had great potential for HPV quality control, as WCID eliminated the mobilization step required by the conventional single-point detection. In the example, the five HPVs displayed pI values of 8.43 ± 0.06 (n = 10; HPV 6L1), 8.70 ± 0.04 (n = 10; HPV 11L1), 7.94 ± 0.05 (n = 18; HPV 16L1), 7.57 ± 0.04 (n = 18; HPV 18L1), and 8.45 ± 0.05 (n = 10; HPV 58L1). This CIEF-WCID platform could be a powerful analytical tool for characterization, process development support, release testing, and stability study in pharmaceutical industry.
采用全柱成像检测（WCID）的毛细管等电聚焦（CIEF）可作为表征和鉴定人乳头瘤病毒（HPV）的有用工具。本文是关于用CIEF-WCID法测定HPV等电点的初步报告。在本研究中，选择用于优化的检测成分包括两性电解质、添加剂、甲基纤维素浓度、HPV浓度、盐浓度和聚焦时间。然后对优化后的CIEF-WCID方法进行了HPV 16L1和HPV 18L1的验证。结果，实现了一种分析HPV等电点值的精确方法，相对标准偏差（RSD）<1.0%。HPV峰型具有可重复性。由于WCID消除了传统单点检测所需的迁移步骤，CIEF-WCID在HPV质量控制方面具有很大潜力。在该实例中，五种HPV的等电点值分别为8.43±0.06（n = 10；HPV 6L1）、8.70±0.04（n = 10；HPV 11L1）、7.94±0.05（n = 18；HPV 16L1）、7.57±0.04（n = 18；HPV 18L1）和8.45±0.05（n = 10；HPV 58L1）。这个CIEF-WCID平台可成为制药行业中用于表征、工艺开发支持、放行检测和稳定性研究的强大分析工具。
