吴飞 赵明娜 郭巧梅 娄加陶△.基于磁性纳米微球的Gamma干扰素酶联免疫检测方法建立[J].,2015,15(18):3410-3414 |
基于磁性纳米微球的Gamma干扰素酶联免疫检测方法建立 |
Establishment of Enzyme-Linked Immunosorbent Assay forInterferon-gamma Detection Based on Immunomagnetic Beads |
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DOI: |
中文关键词: 磁性微球 Gamma干扰素 磁珠酶联免疫分析法 |
英文关键词: Magnetic particle IFN-gamma Immunomagnetic bead ELISA |
基金项目:国家自然科学基金项目(81372193);上海申康医院发展中心临床科室能力建设项目(SHDC22014011);
上海市胸科医院科技发展基金重大重点项目(2014YZDC10100) |
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中文摘要: |
目的:建立及评价使用磁性纳米微球作为固相载体的人酌干扰素(Interferon-gamma,IFN-gamma)双抗体夹心酶联免疫吸附实验
(Enzyme-linked immunosorbent assay,ELISA)检测方法。方法:以杂化细乳液合成法制备磁性纳米微球,将其作为免疫检测的固相
载体。将磁性微球与IFN-酌抗体进行偶联,建立基于磁性微球的ELISA 检测方法,检测人IFN-gamma,绘制IFN-gamma标准曲线并进行方法
学评价。结果:获得包被有人IFN-gamma抗体的免疫微球, 抗体偶联率为54.5 %。用它建立IFN-gamma的双抗体夹心的ELISA 检测方法,检
测范围为0-1000 pg/mL,相关系数为0.9996,灵敏度23.2 pg/mL,功能灵敏度0 pg/mL,批内和批间变异系数(Coefficients of
Variance,CVs)<8 %,检测总共需要2 小时。结论:成功制备了IFN-酌免疫微球并建立了定量检测人IFN-gamma的双抗体夹心磁珠
酶联免疫方法。 |
英文摘要: |
Objective:To establish and evaluate a double Interferon-gamma (IFN-gamma) antibody sandwich Enzyme-linked
immunosorbent assay(ELISA)which was based on immunomagnetic beads for human IFN-gamma detection.Methods:Magnetic nanoparticle
were prepared through a hybrid miniemulsion method and coupled with hunman IFN-Gamma antibody as solid phase. IFN-Gamma Antibody was
coupled to magnetic nanoparticle. ELISA based on immunomagnetic beads for human IFN-Gamma detection was established and evaluated.
Then the standard curve was drawn and assay performance characteristics of the new method was evaluated.Results:Immunoparticles
coated human IFN-gamma antibody were obtained with a coupling efficiency of 54.5 %. Immunoparticles were used to establish the double
IFN-gamma antibody sandwich system with detection range of 0-1000 pg/mL, coefficiency of correlation of 0.9996, sensitivity of 23.2 pg/mL,
functional sensitivity of 0 pg/mL, and intra- and inter-assay coefficients of variation (CVs)<8 %. The total test time was 2 hours.Conclusion:A sandwich ELISA was successfully established for quantitative IFN-gamma detection based on immunomagnetic beads. |
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