| 摘要: |
| [目的]建立一种敏感性高、特异性强的抗苦马豆素(Swainsonine,SW)抗体的ELISA检测方法.[方法]用SW人工抗原(SW-BSA)免疫小鼠,制备抗SW血清,通过ELISA法进行血清抗SW抗体效价检测,比较以SW-OVA或SW为包被抗原的检测结果,分析包被抗原浓度、封闭剂等对测定结果的影响,以确定测定的最佳条件.[结果]以SW-OVA为包被抗原的检测结果明显优于以SW为包被抗原;抗苦马豆素抗体的最佳测定条件为:以1.0 μg/mL SW-OVA作为包被抗原,于4 ℃包被过夜或37 ℃包被2 h,选用15 g/L的明胶作为封闭剂,酶标抗体的工作浓度为1∶1 500.[结论]建立的抗苦马豆素抗体ELISA检测方法,特异性强、稳定性高,为进一步研究苦马豆素人工抗原的免疫原性等奠定了基础. |
| 关键词: 苦马豆素 抗苦马豆素抗体 ELISA检测 |
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| 基金项目:国家自然科学基金项目（30571315）；高等学校博士学科点专项科研基金项目（20030712017） |
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| Establishment of indirect ELISA for detecting the antibody against swainsonine |
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| Abstract: |
| 【Objective】 This paper aimed to establish an indirect ELISA to determine the level of antibody against SW with high sensibility and specificity.【Method】 The mice antiserum was prepared by inoculating the mice with SW-BSA.The antibody against SW was measured through indirect ELISA using SW-OVA or SW as coating antigen and the effects of density of coating antigen and different impacting media on the results were analyzed to determine the optimal conditions of the established indirect ELISA.【Result】 Using the SW-OVA as coating antigen was better than SW.The results showed that the optimal conditions of the method were as follows:the density of coating antigen(SW-OVA) 1.0 μg/mL,the working concentration of enzyme labelled antibody of goat against mouse IgG 1∶1 500,and the impacting medium gelatine of 15 g/L.【Conclusion】 The method was verified with high specificity and stability via the determination of the antibody against SW in serum of mice,thus providing a basic method for further research of SW artificial antigen. |
| Key words: swainsonine antibody against SW ELISA |
