Detection of Influenza A Virus Nucleoprotein Through the Self-Assembly of Nanoparticles in Magnetic Particle Spectroscopy-Based Bioassays: A Method for Rapid, Sensitive, and Wash-free Magnetic Immunoassays

TL;DR

This paper presents a novel magnetic immunoassay method using nanoparticle self-assembly and magnetic particle spectroscopy to rapidly and sensitively detect influenza nucleoprotein without washing steps.

Contribution

It introduces a new biosensing scheme combining MNP self-assembly and MPS for ultralow concentration detection of viral proteins.

Findings

Achieved 44 nM detection sensitivity for H1N1 nucleoprotein.

Demonstrated rapid detection within 10 seconds using MPS.

Proposed multiple MNP self-assembly models to interpret results.

Abstract

Magnetic nanoparticles (MNPs) with proper surface functionalization have been extensively applied as labels for magnetic immunoassays, carriers for controlled drug/gene delivery, tracers and contrasts for magnetic imaging, etc. Here, we introduce a new biosensing scheme based on magnetic particle spectroscopy (MPS) and the self-assembly of MNPs to quantitatively detect H1N1 nucleoprotein molecules. MPS monitors the harmonics of oscillating MNPs as a metric for the freedom of rotational motion, thus indicating the bound states of MNPs. These harmonics can be readily collected from nanogram quantities of iron oxide nanoparticles within 10 s. H1N1 nucleoprotein molecule hosts multiple different epitopes that forms binding sites for many IgG polyclonal antibodies. Anchoring IgG polyclonal antibodies onto MNPs triggers the cross-linking between MNPs and H1N1 nucleoprotein molecules, thereby forming MNP self-assemblies. Using MPS and the self-assembly of MNPs, we achieved the sensitivity of 44 nM (442 pmole) for detecting H1N1 nucleoprotein. In addition, the morphologies and the hydrodynamic sizes of the MNP self-assemblies are characterized to verify the MPS results. Different MNP self-assembly models such as classical cluster, open ring tetramer, chain model as well as multimers (from dimer to pentamer) are proposed in this paper. Herein, we claim the feasibility of using MPS and the self-assembly of MNPs as a new biosensing scheme for detecting ultralow concentrations of target biomolecules, which can be employed as rapid, sensitive, and wash-free magnetic immunoassays.