| 摘要: |
| 【目的】合成并鉴定杠柳新苷T人工抗原，制备多克隆抗体，为进一步开展杠柳新苷类化合物作用机理的研究奠定基础。【方法】以杠柳新苷T为起始反应物，与丁二酸酐反应生成杠柳新苷T的衍生物(半抗原)，采用HPLC-ELSD、MS和NMR对衍生物的结构进行鉴定；将半抗原与载体蛋白（BSA和OVA）进行偶联，采用紫外图谱对人工抗原进行鉴定，并采用2,4,6-三硝基苯磺酸(TNBS)法测定结合比；以乳化的人工抗原免疫8周龄雌性Balb/c小鼠获得杠柳新苷T的多克隆抗体，并测定其效价。【结果】成功合成杠柳新苷T的半抗原和人工抗原，半抗原和载体蛋白（BSA和OVA）的结合比分别为5∶1和7∶1。免疫小鼠后产生针对杠柳新苷T的特异性抗体，其效价为1∶1 600，对杠柳新苷T的半数抑制浓度（IC50）为12.429 4 μg/mL。【结论】制备了杠柳新苷T的多克隆抗体，可以用于杠柳新苷T作用靶标的定位研究。 |
| 关键词: 杠柳新苷T 半抗原 人工抗原 多克隆抗体 |
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| 基金项目:国家公益性行业（农业）科研专项（200903052）；国家自然科学基金项目(30870344,31171868)；中央高校基本科研业务费专项（QN2011058） |
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| Synthesis of artificial antigen and preparation of polyclonal antibody against Periplocoside T |
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| Abstract: |
| 【Objective】This research aimed to synthesize and identify artificial antigen of Periplocoside T and to prepare polyclonal antibody of Periplocoside T.【Method】Periplocoside T derivative (hapten) was synthesized by combining Periplocoside T with succinic anhydride.The product was identified by HPLC-ELSD,MS and NMR.The hapten was conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to form the complete antigens identified by ultraviolet spectrum (UV) through carbodiimide method.【Result】Hapten and artifical antigen of Periplocoside T were synthesized successfully.The ratios of hapten-BSA and hapten-OVA were 5∶1 and 7∶1,respectively.Competitive indirect enzyme linked immunosorbent assay (ELISA) indicated the existence of specific antibodies of Periplocoside T in the antisera.The titer of antisera was 1 600 and the IC50 was 12.429 4 μg/mL.【Conclusion】The polyclonal antibody of Periplocoside T can be applied in future research on insecticide mechanism of Periplocoside T. |
| Key words: Periplocoside T hapten antigen polyclonal antibody |
