School of Pharmacy, Xi'an Jiaotong University, Xi'an 710061, PR China.
School of Pharmacy, Xi'an Jiaotong University, Xi'an 710061, PR China.
Talanta. 2019 Jun 1;198:55-62. doi: 10.1016/j.talanta.2019.01.088. Epub 2019 Jan 25.
Immunoassay as a primary analytical tool is widely applied in the field of analysis. However, biological antibodies used in the routine immunoassay exhibit high cost and poor stability. Herein, in this study, a biomimetic ELISA method using molecularly imprinted polymers as a alternative to antibodies was developed. The molecularly imprinted polymers were prepared through dopamine polymerization using FeO nanoparticles as magnetic nuclei, bovine serum albumin as a template, dopamine as a functional monomer and crosslinking agent. The molecularly imprinted polymers were characterized by scanning electron micrographs, transmission electron microscope, X-ray photoelectron spectroscopy, thermogravimetric analysis, vibrating sample magnetometer and X-ray diffractometer, respectively. The detection range of the established biomimetic ELISA method was 5-1000 μg mL. The method exhibited high selectivity for bovine serum albumin compared with other proteins and good recovery ranging from 89.0% to 102.3% was obtained from spiked bovine serum samples. The results showed that the method by using molecularly imprinted polymers as biomimetic antibodies could be used to detect bovine serum albumin rapidly in bovine serum with a high sensitivity and accuracy. This study demonstrates that the preparation of molecularly imprinted polymers by dopamine polymerization can produce material with high affinity and potential to replace antibodies in biomimetic ELISA for protein detection.
免疫测定作为一种主要的分析工具，广泛应用于分析领域。然而，常规免疫测定中使用的生物抗体成本高且稳定性差。在此，本研究开发了一种使用分子印迹聚合物作为抗体替代物的仿生 ELISA 方法。分子印迹聚合物是通过多巴胺聚合，以 FeO 纳米粒子为磁核、牛血清白蛋白为模板、多巴胺为功能单体和交联剂制备的。通过扫描电子显微镜、透射电子显微镜、X 射线光电子能谱、热重分析、振动样品磁强计和 X 射线衍射仪分别对分子印迹聚合物进行了表征。所建立的仿生 ELISA 方法的检测范围为 5-1000μg/mL。与其他蛋白质相比，该方法对牛血清白蛋白具有较高的选择性，从添加牛血清样品中获得的回收率在 89.0%至 102.3%之间。结果表明，使用分子印迹聚合物作为仿生抗体的方法可以用于快速检测牛血清中的牛血清白蛋白，具有较高的灵敏度和准确性。本研究表明，通过多巴胺聚合制备的分子印迹聚合物可以产生具有高亲和力的材料，有望替代蛋白质检测中的抗体用于仿生 ELISA。
