2013 Fiscal Year Final Research Report

Establishment of maker for early diagnosis of systemic autoimmune diseases based on de-phosphorylation of autoimmogen U1-68k

Research Project

Project/Area Number	24790564
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Multi-year Fund
Research Field	Laboratory medicine
Research Institution	Kinki University
Principal Investigator	NAGAI Kouhei 近畿大学, 生物理工学部, 講師 (70500578)
Project Period (FY)	2012-04-01 – 2014-03-31
Keywords	自己免疫疾患 / 翻訳後修飾 / 診断マーカー / 自己抗体
Research Abstract	In this study, we tried to establish an useful method for detecting a de-phosphorilated form of U1-68k autoimmunogen, which was found to be increased in peripheral blood mononuclear cells of patients of systemic lupus erythematosus or Mixed connective tissue diseases. As a substitution of a conventional 2D-western blot method, we examined three methods; (1) an immunological method, (2) a phos-tag electrophorsis, and (3)an isoelectric focusing electrophoresis-western blot. As a result, we found that the isoelectric focusing electrophoresis-western blot method is most suitable for the detection of the de-phosphorilated form of U1-68k. Moreover, we found that whole cell extract prepared by cell lysis buffer containing SDS can be used for detecting U1-68k. In conclusion, combination of isoelectric focusing electrophoresis-western blot and whole cell extract by SDS-containing cell lysis buffer can shorten analsys time from 5 days to 2.5 days.