Lin C, Gulbis B, Delobbe E, Cotton F, Vertongen F
Department of Clinical Chemistry, Hôpital Erasme, Université Libre de Bruxelles, Brussels, Belgium.
J Chromatogr B Biomed Sci Appl. 1998 Nov 20;719(1-2):47-54. doi: 10.1016/s0378-4347(98)00418-6.
A new separation method of human globin chains by micellar electrokinetic capillary chromatography (MECC) is described. In this method, a 25 mM phosphate buffer (pH 2.5) containing 7 M urea and 1% (w/v) reduced Triton X-100 buffer system was used. All experiments were performed in a 47 cmx50 microm I.D. uncoated fused-silica capillary. The separation voltage was set at 19 kV. Normal globin chains derived from normal adults and newborns, alpha, beta, delta, Ggamma and Agamma globin chains as well as common variant globin chains were successfully separated within 20 min. High reproducible migration times of globin chains (CVs of intra- and inter-assay were less than 1% and 2% respectively), and quantification of Ggamma and Agamma chains (CVs for intra- and inter-assay were less than 5% and 10%, respectively) were obtained. This new MECC method provides primary information on structural modification of globin chains. It can be an important diagnostic tool in clinical laboratory practice in the field of hemoglobinopathies.
描述了一种通过胶束电动毛细管色谱法（MECC）分离人珠蛋白链的新方法。在该方法中，使用了含有7 M尿素和1%（w/v）还原 Triton X - 100的25 mM磷酸盐缓冲液（pH 2.5）缓冲体系。所有实验均在一根47 cm×50 μm内径的未涂层熔融石英毛细管中进行。分离电压设定为19 kV。来自正常成年人和新生儿的正常珠蛋白链、α、β、δ、Gγ和Aγ珠蛋白链以及常见的变异珠蛋白链在20分钟内成功分离。获得了珠蛋白链高度可重复的迁移时间（批内和批间变异系数分别小于1%和2%），以及Gγ和Aγ链的定量结果（批内和批间变异系数分别小于5%和10%）。这种新的MECC方法提供了关于珠蛋白链结构修饰的初步信息。它可以成为血红蛋白病领域临床实验室实践中的一种重要诊断工具。
