Study on the polymorphisms of human serum proteins by using the methods of isoelectric focusing and immunoblotting.

用等电聚焦和免疫印迹的方法研究了人血清蛋白的遗传多态性。对等电聚焦凝胶(聚丙烯酰胺或琼脂糖凝胶)和电极液的厚度和组成进行了修饰，以获得良好的蛋白质条带分离。超薄凝胶(0.1 mm厚)可用于Tf、GC和PI亚型的分型，高浓度凝胶(T=10%，C=3%)可很好地分离PI亚型(如M1M3和M2M3)。神经氨酸酶处理的血清标本等电聚焦时可检测到BF亚型。该凝胶(pH 3~10)含有10%蔗糖和1.5M尿素。表型F分为两个亚型，分别为FA和FB。研究了日本人IgA肾病(IgA-N)和特发性膜性肾病(IMN)患者BF亚型的分布，发现IMN与BF FA(p&lt；0.05)和BF Fb(p&lt；0.01)显著相关。在C7基因多态性研究过程中，发现日本家系成员携带C7沉默等位基因(C7*QO)。C7*QO是单倍型C7*QO-C6*B，C7*QO可能与C6*B相关。在含有尿素的凝胶中，C9蛋白被分成几条带，但没有发现多态性。在含有高浓度尿素的凝胶中，&lt；beta&gt；1H蛋白被分成两组：阳极组和中间组。在中间区域的谱带群中发现了这种多态性，但这些谱带的遗传方式还不完全清楚。应该对阳极区的条带进行分析，以证明&lt；beta&gt；1H的遗传多态。

The genetic polymorphisms of human serum proteins were studied by using the methods of isoelectric focusing and immunoblotting. The thickness and the composition of the isoelectric focusing gel (polyacrylamide or agarose), and electrode solution were modified to obtain the good separation of protein bands. An ultrathin-layer (0.1 mm thick) gel was useful for TF, GC and PI subtypings, and a high concentration gel (T=10%, C=3%) gave good separation of PI subtypes (e.g. M1M3 and M2M3).BF subtypes were detected when neuraminidase-treated serum samples were subjected to isoelectric focusing. The gel (pH 3-10) contained 10% sucrose and 1.5 M urea. The phenotype F was separated into two subtypes, named FA and FB. The distribution of BF subtypes in Japanese patients with IgA nephropathy (IgA-N) or idiopathic membranous nephropathy (IMN) was studied, and the significant association of IMN with BF FA (p<0.05) and that with BF FB (p<0.01) were found.In the course of the study of C7 polymorphism, Japanese families with members carrying C7 silent allele (C7*QO) were found. C6 types in the family members were also examinined; C7*QO was transmitted from a parent to offsprings as a haplotype C7*QO-C6*B. It seems remarkable that C7*QO may be associated with C6*B.The genetic polymorphisms of C9 and <beta>1H were investigated. The C9 proteins were separated into several bands in a gel containing urea, but the polymorphism was not found. The <beta>1H proteins were separated into two groups of bands in a gel containing high concentration of urea; an anodal- and a middle-region groups. The polymorphism was found in the middle region band groups, however, the inheritance of the bands was not fully made clear. The analysis of the bands in the anodal region should be done to demonstrate the genetic polymorphism of <beta>1H.