利奈唑胺单抗的制备及其荧光免疫层析检测法的建立

发布时间：2018-03-09 09:42

本文选题：利奈唑胺　切入点：利奈唑胺衍生物　出处：《药物分析杂志》2017年07期 　论文类型：期刊论文

【摘要】：目的:建立一种简便、快速、特异的利奈唑胺血药浓度荧光免疫层析检测法。方法 :将脱乙酰利奈唑胺定向合成具有单一羧基结构的利奈唑胺衍生物。采用碳二亚胺法将利奈唑胺衍生物与牛血清蛋白和卵清蛋白分别偶联制备完全抗原。将利奈唑胺-牛血清蛋白偶联物免疫Balb/c小鼠制备单克隆抗体。荧光标记利奈唑胺单抗,将利奈唑胺-卵清蛋白完全抗原和羊抗兔IgG抗体喷涂于硝酸纤维素膜分别作为检测线和质控线,以竞争性免疫抑制反应模式建立了利奈唑胺血药浓度荧光免疫层析检测法。基于以上检测方法建立利奈唑胺的标准曲线,并对该检测法进行方法学评价。并采用HPLC方法测定,对两方法的结果进行比较。结果 :通过优化体系,所建立的检测法IC_(50)=5.28μg·mL~(-1),线性范围为0.78~50μg·mL~(-1),灵敏度为0.61μg·mL~(-1),精密度均小于10%,回收率为78.4%~123%。与14种与利奈唑胺结构相似以及联合用药相关药物无明显交叉反应。分别检测67份临床利奈唑胺血样与HPLC法对比,发现2种方法相关性较好,R~2=0.929,大于0.90。结论:本实验成功建立了一种操作简便、快速、高灵敏度、特异性强的利奈唑胺荧光免疫层析检测方法,为进一步临床血药浓度监测的应用打下了基础。
[Abstract]:Objective: to establish a simple and rapid method, A specific fluorescent immunochromatographic assay for the determination of serum concentration of linazolamide. Methods: deacetylleprazolium was used to synthesize linazolamide derivatives with a single carboxyl group. The complete antigen was prepared by coupling bovine serum protein and ovalbumin respectively. The Balb/c mice were immunized with linazolamide and bovine serum protein conjugate to prepare monoclonal antibody. The complete antigen of linazolamine ovalbumin and goat anti-rabbit IgG antibody were sprayed on the nitrocellulose membrane as the detection line and the quality control line, respectively. Based on the competitive immunosuppressive reaction model, a fluorescence immunochromatographic assay for the determination of linazolamine in blood was established. Based on the above methods, the standard curve of linazolamide was established, and the method was evaluated. The method was determined by HPLC method. The results of the two methods are compared. Results: by optimizing the system, The linear range of the method is 0.78 渭 g 路mL ~ (-1), the sensitivity is 0.61 渭 g 路mL ~ (-1), the precision is less than 100.The recovery rate is 78.4m ~ (-1). There is no obvious cross reaction with 14 kinds of drugs similar to the structure of linazolamide and related drugs. The blood samples of nezolamine were compared with HPLC method. It was found that the two methods had a good correlation, which was greater than 0.90.Conclusion: a simple, rapid, sensitive and specific fluorescent immunochromatographic assay was established for the detection of linazolamine. It lays a foundation for the further application of blood drug concentration monitoring.
【作者单位】：湖州市中心医院;中科院湖州营养中心;
【基金】：湖州市科技局公益性技术应用研究(一般)项目,2013GYB12
【分类号】：R392-33

【参考文献】