天然产物莪术醇单克隆抗体的制备

Formation of monoclonal antibody against a major natural product curcumol

关键词:  天然产物  莪术醇  单克隆抗体  制备

Key words:  natural product  curcumol  monoclonal antibody  preparation

摘要:

为制备小分子化合物莪术醇的单克隆抗体，先将莪术醇（curcumol）与载体蛋白牛血清蛋白（BSA）偶联形成完全抗原，用基质辅助激光解吸飞行时间质谱法（MALDI TOF MS）鉴定莪术醇人工抗原的偶联率，然后采用杂交瘤技术获得杂交瘤株，并对其进行小鼠腹水的制备与纯化。结果表明：莪术醇半抗原与载体的偶联比为196，单克隆抗体的腹水效价为1∶51 200，获得的单克隆抗体只与莪术醇抗原发生特异反应，聚丙烯酰胺凝胶电泳((SDS PAGE)显示，单克隆抗体重链的分子量为5 000，轻链的分子量为2 500。得到了莪术醇单克隆抗体，为利用免疫分析技术对莪术类药材进行质量控制与检测奠定了理论基础。

Abstract:

To get monoclonal antibody(mAb)against crude drug curcumol，curcumol carrier protein conjugates were synthesized by the method that curcumol was conjugated with BSA. The ratio of hapten and bovine serum albumin in an antigen conjugate was determined by matrix assisted laser desorption/ionization mass spectrometry〖JP3〗（MALDI TOF MS）. A hybridoma secreting monoclonal antibody against curcumol was produced by hybridoma technology，then preparation and purification of curcumol mAb use mice hybridoma ascitic fluid. Results showed that the ratio of hapten and bovine serum album was 19.6，the ELISA titers in the ascite fluids of the curcumol mAb were 1∶51 200，ELISA analysis also proved that the curcumol mAbs reacted specifically with curcuoml antigen，polyacrylamide gel electrophoresis((SDS PAGE)assay showed that the curcumol heavy chain was 5.0×104 and the light chain was 2.5×104. Conclusion the specific anti curcumol mAbs were prepared，which would lay a theoretical foundation for detection and qulity control and immunoassay of curcuma kwangsiensis in Guangxi producing areas by Immunoassay.