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J Chromatogr A. 2009 Dec 25;1216(52):9028-33. doi: 10.1016/j.chroma.2009.08.027. Epub 2009 Aug 14.
This article provides an overview of a 2D agarose electrophoretic procedure for the characterization of semi-synthetic Haemophilus influenzae type b meningitis vaccines that were prepared for the immunization of small children. The analysis of such vaccines has been particularly challenging because the vaccine particles (i) are highly negatively charged, (ii) are as large as or even larger than intact viruses, and (iii) have a continuous (polydisperse) size distribution because of randomizing steps in the vaccine production (sonification and crosslinking). As a result of these characteristics, 1D electrophoresis of the vaccines produced smears without discernable peaks, but with a second dimension of separation a characteristic vaccine fingerprint was obtained. Whereas O'Farrell gels can accomplish a 2D separation according to size and charge for samples with protein-sized particles, nondenaturing 2D agarose electrophoresis achieves a similar result for much larger virus-sized particles. The separation principle, however, is different. Even though the 2D electrophoretic method was developed from 1983 to 1995, it remains a promising tool for vaccine quality control and for predicting vaccine effectiveness. Modern technology makes the analysis significantly more practical and affordable than it was more than 10 years ago, and the method is applicable to a variety of conjugated vaccines and complex mixtures of virus-sized particles.
本文概述了一种用于鉴定半合成乙型流感嗜血杆菌脑膜炎疫苗的 2D 琼脂糖电泳方法，这些疫苗是为小儿免疫而制备的。由于疫苗颗粒具有以下特点，因此对这些疫苗的分析一直具有挑战性：（i）带高度负电荷，（ii）大小与完整病毒相当，甚至更大，以及（iii）由于疫苗生产中的随机化步骤（超声处理和交联），具有连续（多分散）的粒径分布。结果，1D 电泳产生的疫苗斑点没有可辨别的峰，但通过二维分离获得了特征性的疫苗指纹图谱。尽管 O'Farrell 凝胶可以根据蛋白大小颗粒的大小和电荷对样品进行 2D 分离，但非变性 2D 琼脂糖电泳对于更大的病毒大小颗粒也能达到类似的效果。然而，分离原理是不同的。尽管该 2D 电泳方法是在 1983 年至 1995 年之间开发的，但它仍然是疫苗质量控制和预测疫苗效力的有前途的工具。现代技术使得分析比 10 多年前更加实用和经济实惠，并且该方法适用于各种结合疫苗和病毒大小颗粒的复杂混合物。
