Righetti P G, Gianazza E, Bianchi-Bosisio A, Wajcman H, Cossu G
Department of Biomedical Sciences and Technology, University of Milano, Italy.
Electrophoresis. 1989 Aug-Sep;10(8-9):595-9. doi: 10.1002/elps.1150100811.
The applications of isoelectric focusing in immobilized pH gradients to the analysis of (i) human hemoglobin mutants, (ii) animal hemoglobin mutants (from cattle, sheep, dog and mouse), and (iii) tryptic digests of alpha and beta chains, are discussed and evaluated. Immobilized pH gradients appear to be an excellent tool for screening of genetic polymorphism and for detecting "silent mutants", i.e. those substitutions involving amino acids with nonionizable side chains. At present, not even capillary zone electrophoresis, claimed to have a resolving power equivalent to 1 million theoretical plates, has shown a resolution capability comparable to that of immobilized pH gradients, at least in the field of protein separation.
讨论并评估了等电聚焦在固定化pH梯度中用于分析（i）人类血红蛋白突变体、（ii）动物血红蛋白突变体（来自牛、羊、狗和小鼠）以及（iii）α链和β链的胰蛋白酶消化产物的应用。固定化pH梯度似乎是筛选遗传多态性和检测“沉默突变体”（即涉及具有非离子化侧链氨基酸的取代）的优秀工具。目前，即使是号称具有相当于100万理论塔板数分离能力的毛细管区带电泳，至少在蛋白质分离领域，也未表现出与固定化pH梯度相当的分辨率。
