Tissot J D, Schneider P
Centre de Transfusion sanguine de la Croix-Rouge Suisse, Lausanne.
Rev Fr Transfus Hemobiol. 1989 Oct;32(5):345-56. doi: 10.1016/s1140-4639(89)80022-x.
Two-dimensional gel electrophoresis (2-DGE) associated with a very sensitive silver staining is one of the most powerful technique available for protein analysis. Proteins are separated in the first dimension by isoelectric focusing in a pH gradient, and in the second dimension by polyacrylamide gel electrophoresis. Proteins are thus separated according to their charge and to their size. The high resolution of 2-DGE allows the analysis of complex mixture of proteins with a great reproducibility. More than hundred different proteins can be individualized after 2-DGE of plasma or serum samples. Using this technique, we analysed samples obtained from normal donors and from patients with different gammopathies. Our results confirm that the heavy chains of immunoglobulins are easily detected and that the alpha, gamma and mu chains are clearly differentiated from one another. The typical polyclonal heavy and light chains appear as indiscrete spots after staining of the gels. On the contrary, the monoclonal chains appear as discrete spots. Monoclonal heavy chains have a restricted charge heterogeneity. We also analysed cold agglutinins, partially purified from plasma of three patients showing evidence of chronic hemolysis, a positive direct antiglobulin test of the complement type and anti-I autoantibodies. The 2-DGE have clearly demonstrated that these cold agglutinins were monoclonal IgM, when these later were not detectable by immunofixation electrophoresis. We think that the 2-DGE is a very usefull specialized aid to the analysis of such particular immunohematological problems.
二维凝胶电泳（2-DGE）结合非常灵敏的银染法是目前蛋白质分析中最强大的技术之一。蛋白质在第一维通过在pH梯度中进行等电聚焦分离，在第二维通过聚丙烯酰胺凝胶电泳分离。蛋白质因此根据其电荷和大小进行分离。二维凝胶电泳的高分辨率使得能够对复杂的蛋白质混合物进行分析，且具有很高的重现性。血浆或血清样本经过二维凝胶电泳后，可以区分出一百多种不同的蛋白质。使用这项技术，我们分析了来自正常供体和患有不同丙种球蛋白病患者的样本。我们的结果证实，免疫球蛋白的重链很容易被检测到，并且α、γ和μ链能够清晰地相互区分。凝胶染色后，典型的多克隆重链和轻链呈现为不连续的斑点。相反，单克隆链呈现为离散的斑点。单克隆重链具有有限的电荷异质性。我们还分析了从三名患者血浆中部分纯化得到的冷凝集素，这些患者表现出慢性溶血的迹象、补体型直接抗球蛋白试验阳性以及抗-I自身抗体。二维凝胶电泳清楚地表明，这些冷凝集素是单克隆IgM，而免疫固定电泳无法检测到它们。我们认为二维凝胶电泳对于分析此类特殊的免疫血液学问题是一种非常有用的专门辅助手段。
