中国药理学与毒理学杂志Issue(3)：432-437,6.DOI:10.3867/j.issn.1000-3002.2014.03.021
微囊藻毒素-LR 超灵敏均相免疫分析方法的建立与优化
Development and optimization of ultrasensitive homogenous immunoassay for microcystin-LR
摘要
Abstract
OBJECTIVE Apolyclonalantibody-basedhomogeneouschemiluminescenceimmunoas-say was developed and optimized using AlphaLISA technology for the quantitative detection of microcys-tin-LR(MC-LR)inwatersamples.METHODS Thismethodwasbasedonacompetitivemodelin which an immune complex was formed from the ingegral binding of artificial MC-LR antigen-coated lumi-nescene beads,free MC-LR standards or sa mples,antibody and biotinylated second antibody.Next sensor bead were added that approached the i mmune co mplex through biotin-streptavidin interaction. With the exciting light,the energy was passed from the sensor luminescer before a special emission light could be observed.To opti mize the reaction conditions,working dilutions of polyclonal antibody and bioti-nylated second antibody were assayed while the effect of buffer syste ms and ti me of each reaction were evaluated.RESULTS Maininfluencingfactorsoftheassaywerediscussedasworkingdilutionsofpoly-clonal antibody and biotinylated goat anti rabbit IgG,assay buffer and reacting ti me.After opti mization of reaction conditions,MC-LR AlphaLISA could be finished in 40 min,with a sensitivity of 0.006 μg·L-1 and a dynamic range of 0.006 -5 μg·L-1 .The coefficient of variation was below 10% and average recovery was 1 07.7%.Moreover,the cross reactivity rates of MC-RR and MC-RY to MC-LR were 13.2%and0.91%,respectively.CONCLUSION Thismethodishighlysensitiveandspecific,time-saving and quite suitable for high throughput determination of MC-LR water samples.关键词
微囊藻毒素/微囊藻毒素-LR/化学发光测定法/均相免疫分析Key words
microcystin/microcystin-LR/che milu minescent measurments/ho mogeneous i mmuno-assay分类
医药卫生引用本文复制引用
张艺,黄飚,钮伟民,赵灿培,金坚..微囊藻毒素-LR 超灵敏均相免疫分析方法的建立与优化[J].中国药理学与毒理学杂志,2014,(3):432-437,6.基金项目
The project supported by ″333 high-level personnel training project″of Jiangsu Province （BRA20122209）基金项目江苏省“333高层次人才培养工程” （）
