| 基于EDC/sulfo-NHS的羧基化SPIO表面抗体耦联 |
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| 引用本文： | 刘 琳,张小强,张 宇,浦跃朴,尹立红,刘 卉,徐 靓,杨文倩,徐春雨. 基于EDC/sulfo-NHS的羧基化SPIO表面抗体耦联[J]. 南京医科大学学报(自然科学版), 2013, 0(4): 480-484 |
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| 作者姓名： | 刘 琳 张小强 张 宇 浦跃朴 尹立红 刘 卉 徐 靓 杨文倩 徐春雨 |
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| 作者单位： | 东南大学公共卫生学院,环境医学工程教育部重点实验室,江苏 南京 210009;东南大学公共卫生学院,环境医学工程教育部重点实验室,江苏 南京 210009;江苏省生物材料与器件重点实验室,江苏 南京 210009;东南大学公共卫生学院,环境医学工程教育部重点实验室,江苏 南京 210009;东南大学公共卫生学院,环境医学工程教育部重点实验室,江苏 南京 210009;东南大学公共卫生学院,环境医学工程教育部重点实验室,江苏 南京 210009;东南大学公共卫生学院,环境医学工程教育部重点实验室,江苏 南京 210009;东南大学公共卫生学院,江苏 南京 210009;东南大学公共卫生学院,江苏 南京 210009 |
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| 基金项目： | 江苏省生物材料与器件重点实验室开放基金(2012LBMD05);环境医学工程教育部重点实验室开放基金(2012EME005) |
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| 摘 要： | 目的:研究羧基化超顺磁性氧化铁纳米颗粒(SPIO)纳米粒子与anti-EMMPRIN单克隆抗体耦联的有效方法,考察不同pH条件下进行耦联反应的效率及anti-EMMPRIN-SPIO的粒径水平?方法:用EDC/sulfo-NHS法将anti-EMMPRIN单克隆抗体耦联到羧基化SPIO纳米粒子表面,用动态光散射法检测所得anti-EMMPRIN-SPIO的粒径,并用Bradford法和SDS-PAGE法分析不同酰胺化pH条件下的耦联效率?结果:通过EDC/sulfo-NHS的作用,anti-EMMPRIN单克隆抗体有效地耦联到羧基化SPIO表面?pH7.8时酰胺化的anti-EMMPRIN-SPIO纳米粒子的粒径最小,为63.15 nm?pH6.8和pH7.3条件下酰胺化所得上清中的抗体含量较低,分别为1.943 2 × 10-4 g/L和3.511 1 × 10-4 g/L,SDS-PAGE凝胶图谱上可明显观察到pH6.8?pH7.3和pH7.8条件下所制备样品的谱带分别位于55 000 和25 000?结论:EDC/sulfo-NHS法是anti-EMMPRIN单克隆抗体耦联于羧基化SPIO表面的有效方法?在pH6.8?pH7.3?pH7.8条件下酰胺化耦联效率较高,所制备的anti-EMMPRIN-SPIO粒径较小?pH值对EDC/sulfo-NHS介导的耦联反应的耦联效率及anti-EMMPRIN-SPIO的粒径有较大影响?
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| 关 键 词： | EDC/sulfo-NHS 超顺磁性氧化铁纳米颗粒 EMMPRIN 单克隆抗体 耦联 |
| 收稿时间： | 2012-10-19 |
Conjugation of antibody on carboxyl-SPIO nanoparticles based on EDC/sulfo-NHS |
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| Liu Lin,Zhang Xiaoqiang,Zhang Yu,Pu Yuepu,Yin Lihong,Liu Hui,Xu Liang,Yang Wenqian and Xu Chunyu. Conjugation of antibody on carboxyl-SPIO nanoparticles based on EDC/sulfo-NHS[J]. Acta Universitatis Medicinalis Nanjing, 2013, 0(4): 480-484 |
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| Authors: | Liu Lin Zhang Xiaoqiang Zhang Yu Pu Yuepu Yin Lihong Liu Hui Xu Liang Yang Wenqian Xu Chunyu |
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| Affiliation: | School of Public Health,Key Laboratory of Environmental Medicine Engineering, Ministry of Education, Southeast University, Nanjing 210009;School of Public Health,Key Laboratory of Environmental Medicine Engineering, Ministry of Education, Southeast University, Nanjing 210009;Jiangsu Key Laboratory for Biomaterials and Devices, Southeast University, Nanjing 210009;School of Public Health,Key Laboratory of Environmental Medicine Engineering, Ministry of Education, Southeast University, Nanjing 210009;School of Public Health,Key Laboratory of Environmental Medicine Engineering, Ministry of Education, Southeast University, Nanjing 210009;School of Public Health,Key Laboratory of Environmental Medicine Engineering, Ministry of Education, Southeast University, Nanjing 210009;School of Public Health,Key Laboratory of Environmental Medicine Engineering, Ministry of Education, Southeast University, Nanjing 210009;School of Public Health, Southeast University, Nanjing 210009;School of Public Health, Southeast University, Nanjing 210009 |
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| Abstract: | Objective:To conjugate anti-EMMPRIN monoclonal antibody on the surface of carboxyl-SPIO nanoparticles. The efficiency of superparamagnetic iron oxide (SPIO) conjugated with anti-EMMPRIN monoclonal antibody and the size of anti-EMMPRIN-SPIO were investigated in different pH values. Methods:Anti-EMMPRIN monoclonal antibody conjugated SPIO nanoparticles were performed through a coupling strategy of EDC and sulfo-NHS. Bradford assay and SDS-PAGE analysis were used for measuring the conjugation efficiency. The size of anti-EMMPRIN-SPIO was evaluated by dynamic light scattering method. Results:The smallest diameter of 63.15 nm was obtained when pH 7.8 was used. The lower concentration of antibody in supernatant with pH6.8 and pH7.3 was 1.943 2 × 10-4 g/L and 3.511 1 × 10-4 g/L, respectively. Anti-EMMPRIN mAb was detected in lanes of pH6.8, pH7.3 and pH7.8 which revealed two bands with weight of 55 000 and 25 000. Conclusion:EDC/sulfo-NHS method is an efficient method for conjugating anti-EMMPRIN monoclonal antibody on SPIO nanoparticles. Under the amidation condition with pH value of 6.8, 7.3 and 7.8, higher conjugation efficiency and smaller size of anti-EMMPRIN-SPIO were obtained. The results showed that pH value is a very important factor which influences conjugation efficiency and the size of anti-EMMPRIN-SPIO. |
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| Keywords: | EDC/sulfo-NHS SPIO anti-EMMPRIN monoclonal antibody conjugation |
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