Tissot J D, Hochstrasser D F, Spertini F, Schifferli J A, Schneider P
Centre de Transfusion Sanguine de la Croix-Rouge suisse, Lausanne, Switzerland.
Electrophoresis. 1993 Mar;14(3):227-34. doi: 10.1002/elps.1150140137.
High-resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was used to analyze serum samples and purified immunoglobulins (Ig) obtained from "normal" individuals and from patients diagnosed with monoclonal gammopathies (MG) (n = 47; 5 IgA, 15 IgM, 15 IgG, 4 biclonal IgG, 1 IgD, 7 Bence Jones proteins). Polyclonal and monoclonal heavy (H) chains were located at different restricted gel positions according to their isotype. Monoclonal H chains appeared as sets of spots characterized by charge (pI) and size (M(r)) microheterogeneity. Most of the monoclonal gamma chains were not seen on the gels (12/15). Supplementary polypeptides of 45-48 kDa were detected in serum samples containing monoclonal IgM, but were not seen in MG of other isotypes. However, these polypeptides were not specifically associated with monoclonal IgM because they were also found on protein maps of purified polyclonal IgM. Polyclonal light (L) chains appeared as cloudy bands containing several zones of higher density, whereas monoclonal L chains were usually resolved as single sharp spots. In 6 samples, monoclonal L chains were not seen, and in 9 samples, they appeared as two or more spots, characterized by different pI and/or M(r). In one sample obtained from a patient with a biclonal gammopathy, the L chains were resolved as 4 different spots. Our results confirm that 2-D PAGE is an excellent tool to study Ig. Analysis of the L chain region of the gels was particularly informative. Several monoclonal L chains exhibited heterogeneous two-dimensional spot patterns, suggesting that "subtile" clonal mutations of B-cell lineage and/or posttranslational modifications were involved in their production.
高分辨率二维聚丙烯酰胺凝胶电泳（2-D PAGE）用于分析从“正常”个体以及被诊断为单克隆丙种球蛋白病（MG）的患者（n = 47；5例IgA、15例IgM、15例IgG、4例双克隆IgG、1例IgD、7例本周蛋白）获得的血清样本和纯化的免疫球蛋白（Ig）。多克隆和单克隆重链（H链）根据其同种型位于不同的限定凝胶位置。单克隆H链表现为一组斑点，其特征在于电荷（pI）和大小（M(r)）的微异质性。大多数单克隆γ链在凝胶上未被观察到（12/15）。在含有单克隆IgM的血清样本中检测到45 - 48 kDa的补充多肽，但在其他同种型的MG中未观察到。然而，这些多肽并非与单克隆IgM特异性相关，因为它们也在纯化的多克隆IgM的蛋白质图谱上被发现。多克隆轻链（L链）表现为含有几个较高密度区域的模糊条带，而单克隆L链通常解析为单个清晰的斑点。在6个样本中未观察到单克隆L链，在9个样本中，它们表现为两个或更多斑点，其特征在于不同的pI和/或M(r)。在从一名双克隆丙种球蛋白病患者获得的一个样本中，L链解析为4个不同的斑点。我们的结果证实2-D PAGE是研究Ig的优秀工具。对凝胶L链区域的分析特别有信息量。几种单克隆L链表现出异质的二维斑点模式，表明B细胞系的“细微”克隆突变和/或翻译后修饰参与了它们的产生。
