Bird P S, Seymour G J
Infect Immun. 1987 Mar;55(3):771-7. doi: 10.1128/iai.55.3.771-777.1987.
Monoclonal antibodies (MAbs) against the cell surface antigens of Fusobacterium nucleatum 263 were obtained by fusion of murine myeloma cells (P3-NSI/1-Ag4-1) with the splenocytes of BALB/c mice immunized with whole cells of F. nucleatum 263. Screening was performed using an enzyme-linked immunosorbent assay (ELISA) against the immunizing strain, F. nucleatum 263. Further selection was done using a bacterial panel consisting of Bacteroides, Actinomyces, Streptococcus, Fusobacterium, and Escherichia species. Twelve MAbs were selected on the basis of this screening procedure, seven of which reacted specifically with F. nucleatum 263. Two reacted with F. nucleatum 263 and ATCC 25586, and three reacted with F. nucleatum 263, ATCC 25586, and UQD-003 (a clinical isolate) and also cross-reacted with Fusobacterium russii ATCC 25533. The selected MAbs were then further characterized by absorption experiments with suspensions of intact whole bacterial cells, and the residual binding activity of the supernatants was determined in an ELISA. To determine whether the MAbs reacted with the same or different epitopes, pairs of MAbs were reacted together and independently in a checkerboard manner in an ELISA. The additive or nonadditive nature of the reactivity was determined. A competitive inhibition assay was performed using one labeled and selected unlabeled MAbs. The results of these experiments suggested some epitope sharing among the selected MAbs that reacted with a specific antigen on F. nucleatum and also shared cross-reactive antigens with the three strains of F. nucleatum and F. russii.
通过将小鼠骨髓瘤细胞（P3-NSI/1-Ag4-1）与用具核梭杆菌263全细胞免疫的BALB/c小鼠的脾细胞融合，获得了针对具核梭杆菌263细胞表面抗原的单克隆抗体（MAb）。使用针对免疫菌株具核梭杆菌263的酶联免疫吸附测定（ELISA）进行筛选。使用由拟杆菌属、放线菌属、链球菌属、梭杆菌属和大肠杆菌属组成的一组细菌进一步筛选。基于该筛选程序选择了12种单克隆抗体，其中7种与具核梭杆菌263特异性反应。2种与具核梭杆菌263和ATCC 25586反应，3种与具核梭杆菌263、ATCC 25586和UQD-003（一种临床分离株）反应，并且还与具核梭杆菌ATCC 25533发生交叉反应。然后通过用完整全细菌细胞悬液进行吸收实验对所选单克隆抗体进行进一步表征，并在ELISA中测定上清液的残留结合活性。为了确定单克隆抗体是否与相同或不同的表位反应，将成对的单克隆抗体在ELISA中以棋盘格方式一起和独立反应。确定反应性的加性或非加性性质。使用一种标记的和选择的未标记单克隆抗体进行竞争抑制测定。这些实验结果表明，在所选的与具核梭杆菌上的特定抗原反应并且还与具核梭杆菌和具核梭杆菌的三种菌株共享交叉反应性抗原的单克隆抗体之间存在一些表位共享。
