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目的制备红豆杉属植物中抗癌成分紫杉醇(paclitaxel,TAX)的人工抗原及抗血清,为获取分泌抗紫杉醇抗体的单克隆细胞系及分离单链抗体基因、进而以之提高植物中紫杉醇的量,及建立快速检测TAX的酶联免疫吸附测定(ELISA()法提供技术基础。方法将7-木糖基紫杉醇(7-xylotaxol,7-xyl-TAX)经NaIO4氧化打开糖环,与载体蛋白牛血清白蛋白(BSA)反应偶联,制得半抗原-载体蛋白复合物后,用基质辅助激光解吸飞行时间质谱测定其中结合的半抗原数目;以此抗原免疫BALB/c小鼠,制备抗血清,并通过ELISA法检测其抗体效价和特异性。结果合成的人工抗原TAX-BSA中TAX与BSA的结合比约为4～5∶1;免疫小鼠到特异针对TAX的抗血清,TAX抗体的效价为1∶3200。结论成功地合成了TAX的人工抗原,且该抗原有较好的免疫原性。
Objective To prepare the artificial antigen and antiserum of paclitaxel (TAX), an anticancer component of Taxus, in order to obtain the monoclonal cell line secreting anti-paclitaxel antibody and isolate the single-chain antibody gene, and then increase the amount of paclitaxel in the plant. , and the establishment of rapid detection of TAX enzyme-linked immunosorbent assay (ELISA () method provides the technical basis. Methods 7-xylotaxol (7-xylotaxol, 7-xyl-TAX) by NaIO4 oxidation to open the sugar ring, and carrier proteins After coupling with bovine serum albumin (BSA) to prepare hapten-carrier protein complexes, the number of haptens bound to them was determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; BALB/c mice were immunized with this antigen to prepare antibodies. Serum, and its antibody titer and specificity were detected by ELISA.Results The ratio of binding of TAX to BSA in synthetic TAX-BSA was about 4 to 5:1;Immune mice to antiserum specific to TAX, TAX The titer of the antibody was 1:3200.Conclusion TAX artificial antigen was successfully synthesized and the antigen had better immunogenicity.