Simultaneous express immunoassay of multiple cardiac biomarkers with an automatic platform in human plasma.

C-reactive protein, cystatin C, myoglobin, and D-dimer represent the inflammatory or thromboembolic status of the patient and play important roles in early diagnostics of acute myocardial infarction. Each protein can indicate some health problems, but their simultaneous detection can be crucial for differential diagnostics. The express analysis of these proteins in a small drop of plasma was developed using magnetic beads. The suggested method is based on immunomagnetic extraction of the target analyte from plasma samples and its simultaneous labelling by fluorescent dye. Reaction time was optimized for quantification of cardiac biomarkers in the spike solutions and human plasma samples. In this paper, we developed a one-protein detection technique for each cardiac biomarker and united it to a four-protein facility using an automatic platform. The proposed technique requires only 17 µL of the human plasma and takes 14 min for four-protein measuring. The suggested technique covers concentration difference by more than two orders of magnitude and demonstrates analytical applicability by measurements of human plasma samples of 16 volunteers.

Surface-enhanced Raman spectroscopy in tandem with a gradient electric field from 4-mercaptophenylboronic acid on silver nanoparticles.

The surface-enhanced Raman spectroscopy (SERS) signal of a reporter on silver nanoparticles can be effectively gained by gradient electric field application. The external electric field initiates the dielectrophoresis of nanoparticles and their electrically induced dipole-dipole interaction. Owing to dielectrophoresis, the nanoparticles are concentrated in the area of high electrical field strength. The induced dipole-dipole interaction leads to additional coagulation of nanoparticles and formation of hotspots. Both dielectrophoresis and induced dipole-dipole interaction increase the number of hotspots, which leads to a SERS signal growth. These two mechanisms of SERS signal amplification are explained by the dielectrophoresis and Derjaguin-Landau-Verwey-Overbeek theories. The benefits of the surface-enhanced Raman spectroscopy in tandem with the gradient electric field are experimentally confirmed using a SERS-active reporter, 4-mercaptophenylboronic acid which has a characteristic peak at Raman shift of 1586 cm-1, conjugated to silver nanoparticles of 32, 52, 58, and 74 nm in diameter. The SERS signal gain depends on the silver nanoparticle stability, size, and electric field strength. The limit of detection for 4-mPBA in the system under study can be calculated from the concentration plot and equals to 63 nM. The enhancement factor calculated for SERS in tandem with the gradient electric field can reach 106.Graphical abstract.