Shahram Parvaneh
1,2,3 
, Fatemeh Khademi
2, Gisya Abdi
4, Abdolhamid Alizadeh
4,5, Ali Mostafaie
1,2* 1 Department of Immunology, Faculty of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran
2 Medical Biology Research Center (MBRC), Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran
3 Department of Dermatology and Allergology, Faculty of Medicine, University of Szeged, Szeged, Hungary
4 Academic Centre for Materials and Nanotechnology, AGH University of Science and Technology, al. Adama Mickiewicza 30, 30-059 Krakow, Poland
5 Department of Organic Chemistry, Faculty of Physics and Chemistry, Alzahra University, Tehran, 1993893973, Iran
Abstract
Introduction: Further development of magnetic-based detection techniques could be of significant use in increasing the sensitivity of detection and quantification of hepatitis B virus (HBV) infection. The present work addresses the fabrication and characterization of a new bio-nano composite based on the immobilization of goat anti-HBsAg antibody on modified core-shell magnetic nanoparticles (NPs) by (3-aminopropyl) triethoxysilane (APTES), named Fe3O4@SiO2/NH2, and magnetic NPs modified by chitosan (Fe3O4@CS).
Methods: At the first step, Fe3O4 was modified with the silica and APTES (Fe3O4@SiO2/NH2) and chitosan (Fe3O4@CS) separately. The goat anti-HBsAg antibody was activated by two different protocols: Sodium periodate and EDC-NHS. Then the resulted composites were conjugated with activated goat anti-HBsAg IgG. An external magnet collected Bio-super magnetic NPs (BSMNPs) and the remained solution was analyzed by the Bradford method to check the amount of attached antibody to the surface of BSMNPs.
Results: The findings indicated that activation of antibodies by sodium periodate method 15-17 µg antibody immobilized on 1 mg of super magnetic nanoparticles (SMNPs). However, in the EDC-NHS method, 8-10 µg of antibody was conjugated with 1 mg of SMNPs. The resulting bio-magnetic NPs were applied for interaction with the HBsAg target using enzyme-linked immunosorbent assay (ELISA). About 1 µg antigen attached to 1 mg SMNPs, which demonstrated that the fabricated materials are applicable in the detection scope of HBsAg.
Conclusion: In the present study, we developed new antibody-conjugated magnetic NPs for the detection of HBsAg using an efficient conjugation strategy. The results demonstrated that the binding capacity of Fe3O4@SiO2/NH2 was comparable with commercially available products. Our designed method for conjugating anti-HBsAg antibody to a magnetic nanoparticle opens the way to produce a high capacity of magnetic NPs.