抗Salmonella minnesota R_(595)菌脂多糖杂交瘤的建立及其抗体特性的初步鉴定

将煮沸致死的R_(595)菌体免疫BALB/C小鼠,取其脾细胞在PEG介导下与SP_(2/0)细胞融合,经筛选获得2个阳性孔2D_6、3H_4。将2D_6、3H_4阳性孔细胞克隆,传代最终获得8株能稳定分泌抗R_(595)内毒素单抗的杂交瘤株。经鉴定,杂交瘤细胞染色体数90余条,抗体效价(EL1SA法)10~4～10~5,2D_6B_1,2D_6E_7单抗的Ig类型为IgG_1,3H_42F_7、3H_42H_3单抗Ig类型为IgG2_b,轻链均为K链。类脂A抑制试验显示,单抗识别的表位为类脂A或KDO,表明抗体识别内毒素保守区域。间接EL1SA试验表明,抗体能与多种GNB交叉反应,提示抗体具广谱交叉反应性。玻片凝集试验显示,抗体仅能凝集R_(595)菌,不能凝集其它GNB菌。

ESTABLISHMENT OF HYBRIDOMA CELL LINES SECRCTING MONOCLONAL ANTIBODIES AGAINST LIPOPOLYSACCHARIDE OF SALMONELLA MINNESOTA R_(595)

Several hybridoma cell lines secreting monoclonal antibodies (McAbs) against endo-toxin of R596 were established by the fusion of SP2/0 myeloma cells with spleen cells from BALB/C mice immunized with heat -killed whole R595 cells The average number of the chromosomes of the hybridoma cells was more than 90 and the liters of the McAbs in the ascites ranged from 10000 to 100000 with ELISA. The immunoglobulin subclass of McAbs 2D6B, and 2D6E7 was IgG: and that of McAbs 3H42F7 and 3H42H3 was IgG2b- All of their light chains were K chain. The epitopes of R595 recognized by McAbs might be in the conserved lipid Aor KDO, suggested by the results of competitive binding test. The results of the indirect ELISA suggested that McAbs could cross -react to various gram -negative bacilli, which indicated their wide cross - reactivities. But the result of the slide agglutination test showed that McAbs aggluted only R595 and could not agglute others.