Improvements of magnetic nanoparticle assays for SARS-CoV-2 detection using a mimic virus approach

Abstract

Immunoassays with magnetic nanoparticles (MNPs) as markers are a promising approach for the fast and sensitive virus detection. Upon binding of antibody-functionalized MNP on virus proteins, the hydrodynamic diameter increases and a change in the Brownian relaxation time can be measured. In this study, we detect the whole SARS-CoV-2 by mimicking it with streptavidin-coated polystyrene beads with biotinylated spike proteins. Changes of the MNP dynamics are measured by alternating current susceptometry and magnetic particle spectroscopy. Due to the multiple binding sites of MNP and virus, crosslinking enlarges the change of the hydrodynamic diameter. In order to improve the sensitivity and the limit of detection of the assay, the ratio of the virus to the MNP amount R_MV/MNP is investigated in detail. High R_MV/MNP ratios lead to a saturation of the MNPs with viruses, so that the cluster size and therefore the sensitivity decrease again. Additionally, it is found that the smallest virus concentrations can be detected for small MNP concentrations. It is also shown that the R_MV/MNP range that can be used for an unambiguous detection of viruses depends on the virus/MNP concentration; it shifts with increasing MNP concentration to smaller R_MV/MNP values. For very small virus concentrations, an increase of the Brownian relaxation time is detected implying a decrease of the hydrodynamic diameter. Furthermore, the optimal antibody concentration for MNP functionalization was determined. It is also found that a washing process with a centrifuge improves the sensitivity by reliably removing unbound antibodies and eliminating small MNPs with improper functionalization.