岩沙海葵毒素单克隆抗体的制备及其鉴定

目的建立岩沙海葵毒素（Palytoxin，PTX）高特异性单克隆抗体杂交瘤细胞株。方法将PTX偶联成具有免疫功能的PTX-PDP-KLH完全抗原，免疫雌性6—8周龄Balb／c小鼠，利用传统杂交瘤技术制备PTX单克隆抗体。并通过间接酶联免疫吸附测定（ELISA）方法筛选克隆阳性杂交瘤细胞株。结果获得能稳定分泌抗PTX单克隆抗体的杂交瘤细胞株B8，与载体蛋白没有发生交叉反应。SDS—PAGE分析表明，McAb B8在50KD和25KD处各出现一条条带，与IgG的重链和轻链的大小相符。结论研究制备了特异性单克隆抗体PTX，为进一步建立快速检测、鉴定海产品中岩沙海葵毒素的方法，奠定有效的实验基础。

Preparation and identification of monoclonal antibody against palytoxin

Objective To establish monoclonal antibodies against palytoxin. Methods Palytoxin was coupled to KLH and the artificial antigens synthesized were used to immunize Balb/c mice. By fusing murine NS-1 cells with spleen cells from mice immunized obtain a hybridoma cell line steadily secreting monoclonal antibodies against Palytoxin. Furthermore, by indirect ELISA, The clones which could secrete specific monoclonal antibodies were secreened. Results The monoclonal antibodies from B8 showed highly significant reactivity. Conclusion It was feasible to obtain hybridoma cell lines which could secrete monoclonal antibodies against the Palytoxin by hybridoma technology. The sensitivity of the indirect ELISA reached at ng level, the result of which is reliable. The method can be employed to detectPalytoxin in seafood.