Magnetic Relaxation Switching Assay Using IFNα-2b-Conjugated Superparamagnetic Nanoparticles for Anti-Interferon Antibody Detection
Boris Nikolaev,
Ludmila Yakovleva,
Viacheslav Fedorov,
Natalia Yudintceva,
Vyacheslav Ryzhov,
Yaroslav Marchenko,
Alexander Ischenko,
Alexander Zhakhov,
Anatoliy Dobrodumov,
Stephanie E. Combs,
Huile Gao,
Maxim Shevtsov
Affiliations
Boris Nikolaev
Laboratory of Biomedical Nanotechnologies, Institute of Cytology of the Russian Academy of Sciences (RAS), Tikhoretsky Ave., 4, 194064 St. Petersburg, Russia
Ludmila Yakovleva
Laboratory of Biomedical Nanotechnologies, Institute of Cytology of the Russian Academy of Sciences (RAS), Tikhoretsky Ave., 4, 194064 St. Petersburg, Russia
Viacheslav Fedorov
Laboratory of Biomedical Nanotechnologies, Institute of Cytology of the Russian Academy of Sciences (RAS), Tikhoretsky Ave., 4, 194064 St. Petersburg, Russia
Natalia Yudintceva
Laboratory of Biomedical Nanotechnologies, Institute of Cytology of the Russian Academy of Sciences (RAS), Tikhoretsky Ave., 4, 194064 St. Petersburg, Russia
Vyacheslav Ryzhov
Petersburg Nuclear Physics Institute, National Research Centre “Kurchatov Institute”, 188300 Gatchina, Russia
Yaroslav Marchenko
Petersburg Nuclear Physics Institute, National Research Centre “Kurchatov Institute”, 188300 Gatchina, Russia
Alexander Ischenko
Laboratory of Hybridoma Technologies, Saint-Petersburg Pasteur Institute, Mira Str. 14, 197101 St. Petersburg, Russia
Alexander Zhakhov
Laboratory of Hybridoma Technologies, Saint-Petersburg Pasteur Institute, Mira Str. 14, 197101 St. Petersburg, Russia
Anatoliy Dobrodumov
Department of Nuclear Magnetic Resonance, Institute of Macromolecular Compounds of the Russian Academy of Sciences (RAS), Bolshoi pr. 31, 199004 St. Petersburg, Russia
Stephanie E. Combs
Department of Radiation Oncology, Technishe Universität München (TUM), Klinikum Rechts der Isar, Ismaninger Str. 22, 81675 Munich, Germany
Huile Gao
Key Laboratory of Drug-Targeting and Drug Delivery System of the Education Ministry, West China School of Pharmacy, Sichuan University, Chengdu 610041, China
Maxim Shevtsov
Laboratory of Biomedical Nanotechnologies, Institute of Cytology of the Russian Academy of Sciences (RAS), Tikhoretsky Ave., 4, 194064 St. Petersburg, Russia
Type I interferons, particularly IFNα-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 IFNα-2b or anti-IFNα-2b antibodies is of high importance to improve the diagnosis of various pathologies associated with the IFNα-2b disbalance. For evaluation of the anti-IFNα-2b antibody level, we have synthesized superparamagnetic iron oxide nanoparticles (SPIONs) coupled with the recombinant human IFNα-2b protein (SPIONs@IFNα-2b). Employing a magnetic relaxation switching assay (MRSw)-based nanosensor, we detected picomolar concentrations (0.36 pg/mL) of anti-INFα-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@IFNα-2b nanoparticles with the anti-INFα-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@IFNα-2b nanoparticles represents an alternative immunological probe for the estimation of anti-IFNα-2b antibodies that could be further employed in clinical studies.
