Abstract
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−1 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−1 to 6 mg ml−1 with the limit of detection of 60 pg ml−1. This method was successfully used to detect myoglobin in spiked fetal calf serum, with a recovery rate of more than 93%.
Highlights
A biomarker is a characteristic that is accurately measured and evaluated as an indicator of normal biological processes, pathogenic processes, or pharmacologic responses to a therapeutic intervention [1]
The assay uses an excess amount of F-molecularly imprinted polymer (MIP) as a capture molecule, to bind to all of the myoglobin within the sample
The MIP left in the sample was removed and analyzed using fluorescent spectroscopy, with fluorescent intensity directly related to the amount of myoglobin in the sample
Summary
A biomarker is a characteristic that is accurately measured and evaluated as an indicator of normal biological processes, pathogenic processes, or pharmacologic responses to a therapeutic intervention [1]. Myoglobin is the earliest biomarker to appear, following acute myocardial infarction (AMI) [8]. It is an important oxygen carrying monomeric heme-protein that is found in heart and skeletal muscles, with its main function being the uptake of oxygen from oxyhemoglobin in the bloodstream [9]. Mb that is determined in serum can be used as an important biomarker for cardiac injury [10]. The concentration of Mb within serum can quickly become elevated in as little as one hour after AMI and this correlates with the degree of myocardial injury [12, 13]. Current clinical practises use immunoassays, for the detection of AMI biomarkers in blood serum, which have relatively low accuracy and sensitivity, making the monitoring of disease treatment difficult. There is, scope for the development of new and more accurate assays and diagnostics
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
