Wijnen P A, van Dieijen-Visser M P
Department of Clinical Chemistry, Academic Hospital, Maastricht, The Netherlands.
Eur J Clin Chem Clin Biochem. 1996 Jul;34(7):535-45. doi: 10.1515/cclm.1996.34.7.535.
Conditions of serum protein analysis by capillary electrophoresis were optimized and within day, between day and between capillary variations were examined for both migration times and relative peak areas. For the five currently accepted zones, albumin, alpha 1, alpha 2, beta and gamma-globulin, reproducibilities of migration times were in the range of 2.3-3.1% (n = 200 measurements). Although variations in relative peak areas were slightly higher than those obtained by conventional agarose gel electrophoresis, from a resolution perspective, capillary electropherograms provided better detail than the densitometric scans of agarose gel electrophoresis. Precise localization of C3 and transferrin in capillary electrophoresis resulted in more accurate detection of the beta-globulin fraction. When C3 appeared in the gamma-fraction it was not detected as a separate peak in agarose gel electrophoresis, whereas it was in capillary electrophoresis. In artificially prepared mixtures of highly purified albumin and gamma-globulin preparations, best correspondence with theoretical values was found with capillary electrophoresis. Inter-individual variations and reference values were obtained by measuring 140 samples from healthy controls (59 females, 81 males) with both techniques. For capillary electrophoresis the inter-individual variations of the albumin, alpha 1, alpha 2, beta and gamma fractions were respectively 6, 21, 19, 14 and 18% and for agarose gel electrophoresis 5, 20, 17, 18 and 22%. From these results it can be concluded that the more precise localization of the beta- and gamma-globulin fraction results in about 4% lower inter-individual variations in capillary electrophoresis compared to agarose gel electrophoresis. For the other fractions, comparable variations were obtained. Differences between males and females were not significant. For patient samples, a good correlation was found between capillary electrophoresis and agarose gel electrophoresis data for all five protein fractions. We conclude that separation efficiency of capillary electrophoresis is better than that of agarose gel electrophoresis and even weak monoclonal components can easily be distinguished with the capillary electropherogram. Capillary electrophoresis is a qualitatively good, cheap, fast and easy to perform alternative to agarose gel electrophoresis.
优化了毛细管电泳分析血清蛋白的条件，并检测了日内、日间以及不同毛细管之间迁移时间和相对峰面积的变化。对于目前公认的五个区带，即白蛋白、α1、α2、β和γ球蛋白，迁移时间的重现性在2.3 - 3.1%范围内（n = 200次测量）。虽然相对峰面积的变化略高于传统琼脂糖凝胶电泳，但从分辨率角度来看，毛细管电泳图谱比琼脂糖凝胶电泳的光密度扫描提供了更详细的信息。毛细管电泳中C3和转铁蛋白的精确定位使得β球蛋白组分的检测更加准确。当C3出现在γ组分中时，在琼脂糖凝胶电泳中它不会作为一个单独的峰被检测到，而在毛细管电泳中则可以。在人工制备的高纯度白蛋白和γ球蛋白制剂的混合物中，毛细管电泳与理论值的对应性最佳。通过两种技术对140例健康对照样本（59名女性，81名男性）进行测量，获得了个体间差异和参考值。对于毛细管电泳，白蛋白、α1、α2、β和γ组分的个体间差异分别为6%、21%、19%、14%和18%，对于琼脂糖凝胶电泳分别为5%、20%、17%、18%和22%。从这些结果可以得出结论，与琼脂糖凝胶电泳相比，毛细管电泳中β和γ球蛋白组分的更精确定位使得个体间差异降低约4%。对于其他组分，获得了可比的差异。男性和女性之间的差异不显著。对于患者样本，所有五个蛋白组分的毛细管电泳和琼脂糖凝胶电泳数据之间发现了良好的相关性。我们得出结论，毛细管电泳的分离效率优于琼脂糖凝胶电泳，甚至弱单克隆成分也可以通过毛细管电泳图谱轻松区分。毛细管电泳是一种质量好、价格便宜、快速且易于操作的替代琼脂糖凝胶电泳的方法。
