B型肉毒毒素单克隆抗体杂交瘤株的建立

PREPARATION AND CHARACTERIZATION OF MONOCLONAL ANTIBODY AGAINST BOTULINUM NEUROTOXIN TYPE B

[目的]制备、筛选稳定分泌B型肉毒毒素单抗的杂交瘤克隆,为相关研究提供工具。[方法]采用纯化的B型肉毒毒素免疫BALB/c小鼠,用免疫小鼠的脾细胞与骨髓瘤细胞Sp2/0融合,筛选阳性克隆。[结果]得到10株稳定分泌B型肉毒毒素单抗的杂交瘤克隆,1株为IgM,其余9株分泌IgG,亚类为IgG2a。10株单克隆抗体均与B型肉毒毒素呈特异性反应,而与标准A型肉毒毒素无交叉反应,腹水单抗的效价均在105以上。[结论]10株杂交瘤细胞株均能分泌抗B型肉毒毒素单克隆抗体,为B型肉毒毒素的检测和研究提供了工具。

[Objective] To prepare monoclonal antibodies against Botulinum neurotoxin type B as the tool for ralative research. [Methods] BALB/c mice were intraperitoneally immunized with purified Botulinum neurotoxin type B; the splenocytes from immunized mice were fused with myeloma cells Sp2/0. Hybridoma cells were screened by indirect ELISA and limiting dilution method. [ Results] Ten hybridomas producing the McAbs against Botulinum neurotoxin type B were successfully established, one of which was IgM, the others were IgG, and subclasses of them were IgG2a. The titer of ascitic McAbs were all above 1×10^5. [ Conclusion] Ten McAbs against Botulinum neurotoxin type B with high titers are obtained successfully, which will be useful for the detection and ralative research work of Botulinum neurotoxin type B.