Magnetic Relaxation Switching Assay Using IFNα-2b-Conjugated Superparamagnetic Nanoparticles for Anti-Interferon Antibody Detection

Type I interferons, particularly IFNa-2b, play essential roles in eliciting adaptive and innate immune responses, being implicated in the pathogenesis of various diseases, including cancer, and autoimmune and infectious diseases. Therefore, the development of a highly sensitive platform for analysis of either IFNa-2b or anti-IFNa-2b antibodies is of high importance to improve the diagnosis of various pathologies associated with the IFNa-2b disbalance. For evaluation of the anti-IFNa-2b antibody level, we have synthesized superparamagnetic iron oxide nanoparticles (SPIONs) coupled with the recombinant human IFNa-2b protein (SPIONs@IFNa-2b). Employing a magnetic relaxation switching assay (MRSw)-based nanosensor, we detected picomolar concentrations (0.36 pg/mL) of anti-INFa-2b antibodies. The high sensitivity of the real-time antibodies' detection was ensured by the specificity of immune responses and the maintenance of resonance conditions for water spins by choosing a high-frequency filling of short radio-frequency pulses of the generator. The formation of a complex of the SPIONs@IFNa-2b nanoparticles with the anti-INFa-2b antibodies led to a cascade process of the formation of nanoparticle clusters, which was further enhanced by exposure to a strong (7.1 T) homogenous magnetic field. Obtained magnetic conjugates exhibited high negative MR contrast-enhancing properties (as shown by NMR studies) that were also preserved when particles were administered in vivo. Thus, we observed a 1.2-fold decrease of the T2 relaxation time in the liver following administration of magnetic conjugates as compared to the control. In conclusion, the developed MRSw assay based on SPIONs@IFNa-2b nanoparticles represents an alternative immunological probe for the estimation of anti-IFNa-2b antibodies that could be further employed in clinical studies.

Automated summary

Type I interferons, particularly IFNa-2b, play crucial roles in immune responses and are associated with the development of various diseases. Therefore, it is essential to develop a highly sensitive platform for the analysis of IFNa-2b and anti-IFNa-2b antibodies to improve diagnosis.