Research Centre for Smart Materials, Department of Chemistry, School of Natural Sciences, University of Central Lancashire, Preston, PR1 2HE, United Kingdom.
Leicester School of Pharmacy, De Montford University, The Gateway, Leicester, LE1 9BH, United Kingdom.
Nanotechnology. 2021 Feb 26;32(9):095502. doi: 10.1088/1361-6528/abce2d.
We have developed a low-cost molecularly imprinted polymer (MIP)-based fluorometric assay to directly quantify myoglobin in a biological sample. The assay uses a previously unreported method for the development of microwave-assisted rapid synthesis of aldehyde functionalized magnetic nanoparticles, in just 20 min. The aldehyde functionalized nanoparticles have an average size of 7.5 nm ± 1.8 and saturation magnetizations of 31.8 emu g with near-closed magnetization loops, confirming their superparamagnetic properties. We have subsequently shown that protein tethering was possible to the aldehyde particles, with 0.25 ± 0.013 mg of myoglobin adsorbed to 20 mg of the nanomaterial. Myoglobin-specific fluorescently tagged MIP (F-MIP) particles were synthesized and used within the assay to capture myoglobin from a test sample. Excess F-MIP was removed from the sample using protein functionalized magnetic nanoparticles (Mb-SPION), with the remaining sample analyzed using fluorescence spectroscopy. The obtained calibration plot of myoglobin showed a linear correlation ranging from 60 pg ml to 6 mg ml with the limit of detection of 60 pg ml. This method was successfully used to detect myoglobin in spiked fetal calf serum, with a recovery rate of more than 93%.
我们开发了一种基于分子印迹聚合物（MIP）的低成本荧光分析方法，可直接定量生物样品中的肌红蛋白。该方法采用了一种以前未报道的方法，用于开发微波辅助快速合成醛功能化磁性纳米粒子，仅需 20 分钟。醛功能化纳米粒子的平均粒径为 7.5nm±1.8，饱和磁化强度为 31.8emu/g，具有近乎闭合的磁化回线，证实了其超顺磁性。随后我们证明了蛋白质可以与醛基颗粒结合，20mg 纳米材料可吸附 0.25±0.013mg 的肌红蛋白。合成了肌红蛋白特异性荧光标记的分子印迹聚合物（F-MIP）颗粒，并在该分析方法中用于从测试样品中捕获肌红蛋白。使用蛋白质功能化的磁性纳米颗粒（Mb-SPION）从样品中去除多余的 F-MIP，然后使用荧光光谱法分析剩余的样品。获得的肌红蛋白校准曲线显示，线性相关范围为 60pg/ml 至 6mg/ml，检测限为 60pg/ml。该方法成功地用于检测胎牛血清中的肌红蛋白，回收率超过 93%。
