Efficient conjugation of anti-HBsAg antibody to modified core-shell magnetic nanoparticles (Fe3O4@SiO2/NH2)

Shahram Parvaneh; Fatemeh Khademi; Gisya Abdi; Abdolhamid Alizadeh; Ali Mostafaie

doi:10.34172/bi.2021.34

Efficient conjugation of anti-HBsAg antibody to modified core-shell magnetic nanoparticles (Fe₃O₄@SiO₂/NH₂)

Bioimpacts. 2021;11(4):237-244. doi: 10.34172/bi.2021.34. Epub 2021 Apr 28.

Authors

Shahram Parvaneh^{1

2

3}, Fatemeh Khademi², Gisya Abdi⁴, Abdolhamid Alizadeh^{4

5}, Ali Mostafaie^{1

2}

Affiliations

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.

Keywords: Antibody conjugation; Detectiion; HBsAg; Magnetic nanoparticles.