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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,McAbB8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。
Objective To establish a hybridoma cell line with highly specific monoclonal antibody against Palytoxin (PTX). Methods The PTX-PDP-KLH complete antigen with immunopotency was conjugated to immunized Balb / c mice of 6-8 weeks of age. The monoclonal antibody against PTX was prepared by indirect immunofluorescence assay and indirect ELISA (ELISA) screening of clonal positive hybridoma cell lines. Results The hybridoma cell line B8, which can stably secrete anti-PTX monoclonal antibody, was obtained and did not cross-react with the carrier protein. SDS-PAGE analysis showed that McAbB8 showed a band at 50KD and 25KD each, which was consistent with the size of IgG’s heavy chain and light chain. Conclusion The specific monoclonal antibody (PTX) was prepared and studied in this study, which will lay the foundation for the further establishment of a rapid method for the detection and identification of rock and sea anemone toxin in seafood.