Sato H, Yamazaki Y, Kodaira A, Saito H, Maehara N
Department of Veterinary Microbiology, School of Veterinary Medicine and Animal Sciences, Kitasato University, Aomori, Japan.
Zentralbl Veterinarmed B. 1999 Mar;46(2):85-92. doi: 10.1111/j.0931-1793.1999.00209.x.
Thirteen mouse monoclonal antibodies (Mabs) against the protective protein antigen (P64) of Erysipelothrix rhusiopathiae were prepared and partially characterized. The titres of the Mabs varied from 200 to 1,638,400 as determined by enzyme-linked immunosorbent assay (ELISA). Of the 13 Mabs 10, two and one belonged to the IgG2a, IgG1 and IgM subclasses, respectively. All Mabs reacted strongly with the 64 kDa protein and weakly with the 43 kDa protein upon Western blotting of the alkaline extract (AE) of E. rhusiopathiae. The protective activity (PD50/ml) of the 13 Mabs against E. rhusiopathiae infection in mice varied from < 50 to > 50,000. These Mabs were classified into three groups, highly protective Mabs, moderately protective Mabs and Mabs which did not possess protective activity, based on the protective index (ratio of the PD50/ml to the antibody titre). These results suggest that the 64 kDa protein is an effective protective antigen, which is easily cleaved into many small proteins, including the 43 kDa protein, and possesses at least two epitopes related to its protective activity and at least one epitope which is not related to protection of mice against E. rhusiopathiae infection.
制备了13种针对猪丹毒杆菌保护性蛋白抗原（P64）的小鼠单克隆抗体（Mab），并对其进行了部分特性鉴定。通过酶联免疫吸附测定（ELISA）确定，这些单克隆抗体的效价在200至1,638,400之间。13种单克隆抗体中，分别有10种、2种和1种属于IgG2a、IgG1和IgM亚类。在对猪丹毒杆菌碱性提取物（AE）进行蛋白质印迹分析时，所有单克隆抗体均与64 kDa蛋白强烈反应，而与43 kDa蛋白反应较弱。13种单克隆抗体对小鼠猪丹毒杆菌感染的保护活性（PD50/ml）在<50至>50,000之间。根据保护指数（PD50/ml与抗体效价之比），这些单克隆抗体被分为三组：高保护性单克隆抗体、中等保护性单克隆抗体和不具有保护活性的单克隆抗体。这些结果表明，64 kDa蛋白是一种有效的保护性抗原，它很容易被切割成许多小蛋白，包括43 kDa蛋白，并且具有至少两个与其保护活性相关的表位以及至少一个与保护小鼠免受猪丹毒杆菌感染无关的表位。
