Righetti Pier Giorgio
Politecnico di Milano, Department of Chemistry, Materials and Engineering Chemistry Giulio Natta, Milano, Italy.
Electrophoresis. 2006 Mar;27(5-6):923-38. doi: 10.1002/elps.200500525.
The birth and evolution of IEF in conventional carrier ampholyte buffers is reviewed here, from a shaky start during World War II, via desperate attempts of Svensson to create pH gradients by stationary electrolysis of salts, to the development of the IEF theory and the solution of the steady-state equation. The remarkable synthetic process of Ampholines, as ingeniously devised by Vesterberg, is additionally recalled, with a thorough description of the fundamental properties of these amphoteric buffers, creating and maintaining the pH gradient under strong electric fields. The review ends with a mention of the major contributions of B. J. Radola to this field, namely analytical and preparative IEF in granulated Sephadex layers and the development of ultrathin IEF, in polyacrylamide gels as thin as 20-100 mum. The latter technique paved the way to DNA sequencing gels and to CZE. The symptoms of decay are here presented through the simulations of Mosher and Thormann, clearly indicating an isotachophoretic mechanism for pH gradient decay with time. The decay of IEF was the birth of IPGs.
本文回顾了等电聚焦（IEF）在传统载体两性电解质缓冲液中的诞生与发展历程，从二战期间的艰难起步，斯文森通过盐的静态电解来创建pH梯度的绝望尝试，到IEF理论的发展以及稳态方程的求解。此外，还回顾了韦斯特伯格巧妙设计的两性电解质的非凡合成过程，并全面描述了这些两性缓冲剂的基本特性，即在强电场下创建和维持pH梯度。本文最后提到了B. J. 拉多拉对该领域的主要贡献，即在粒状葡聚糖凝胶层中的分析型和制备型IEF以及超薄IEF的发展，后者可用于制备厚度仅为20 - 100微米的聚丙烯酰胺凝胶。后一种技术为DNA测序凝胶和毛细管区带电泳（CZE）铺平了道路。莫舍和托曼的模拟展示了IEF衰退的症状，清楚地表明了pH梯度随时间衰减的等速电泳机制。IEF的衰退催生了固定化pH梯度（IPG）。
