Masuzawa T, Matsumoto T, Nakamura R, Suzuki R, Shimizu T, Yanagihara Y
Department of Microbiology, School of Pharmaceutical Sciences, University of Shizuoka, Japan.
J Gen Microbiol. 1990 Feb;136(2):327-30. doi: 10.1099/00221287-136-2-327.
A protective glycolipid antigen (PAg) was extracted from Leptospira interrogans serovar canicola with chloroform/methanol/water (1:2:0.8, by vol.) and partially purified by silica gel column chromatography. The PAg elicited a protective response in hamsters and in cyclophosphamide-treated mice subsequently challenged with homologous Leptospira. The PAg band was detected as a single smear-like band, corresponding to a protein of 23-30 kDa, by silver-staining in SDS-PAGE. In immunoblots, this band reacted with a monoclonal antibody, A5, which agglutinated serovar canicola and recognized a serovar-specific antigen. Furthermore, the PAg did not migrate on silica gel TLC, but was detected at the origin as a ninhydrin- and naphthol-positive spot. This suggests that PAg is a hydrophilic molecule with a carbohydrate chain that contains amino groups, possibly as amino sugars.
用氯仿/甲醇/水（体积比1:2:0.8）从犬钩端螺旋体血清型犬型中提取一种保护性糖脂抗原（PAg），并通过硅胶柱色谱法进行部分纯化。该PAg在仓鼠和随后用同源钩端螺旋体攻击的环磷酰胺处理小鼠中引发了保护性反应。在SDS-PAGE中，通过银染检测到PAg条带为单一的涂片样条带，对应于23-30 kDa的蛋白质。在免疫印迹中，该条带与单克隆抗体A5反应，A5凝集犬型血清型并识别血清型特异性抗原。此外，PAg在硅胶TLC上不迁移，但在原点处被检测为茚三酮和萘酚阳性斑点。这表明PAg是一种具有碳水化合物链的亲水分子，该碳水化合物链含有氨基，可能是氨基糖。
