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

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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.