摘要
目的:利用小鼠神经生长因子(mNGF)和纯化的兔抗mNGF IgG,建立mNGF直接竞争酶联免疫检测方法。方法:以亲和纯化兔抗mNGF IgG作为包被抗体,封闭、洗涤后加入用稀释液稀释的不同浓度的标准mNGF或样品液和生物素标记的mNGF混合液,于37℃孵育80 min或室温孵育120 min,洗涤后加入辣根过氧化物酶标记的链酶亲和素,洗涤后加入显色液显色,测定D450nm值,通过标准曲线计算待测样品中mNGF含量。结果与结论:建立了mNGF直接竞争酶联免疫检测方法;该方法的灵敏度约20 ng,变异系数为批内≤10%、批间≤15%,回收率为85%～115%;利用该方法检测了3批样品中mNGF的含量。
Objective： Using biotinylated mouse nerve growth factor（mNGF） and rabbit-anti-mNGF IgG to estab-lish direct competition ELISA method for detection of mNGF.Methods： The microwell plates were coated with affinity purified rabbit-anti-mNGF IgG,after blocking and washing,series concentrations of mNGF standard dilu-tions or samples and biotinylated mNGF mixtures were added into the wells.The plates were incubated at 37℃ for 80 min or at room temperature for 120 min,after washing the horseradish peroxidase conjugated streptavidin were added,and then color were developed to detection the D 450nm values.By standard curve,the concentrations of mNGF in samples were calculated.Results Conclusion： The direct competition ELISA method for mNGF was successfully established.The detection sensitivity is 20 ng,the coefficient variation within group is ≤10% and co-efficient variation between groups is ≤15%.The recovery rate of the method is 85%～115%.Using the established method,the contents of mNGF in three batches of samples were evaluated.
出处
《生物技术通讯》
CAS
2011年第5期705-708,共4页
Letters in Biotechnology
