抗hbsag抗体与修饰核壳磁性纳米颗粒(Fe3O4@SiO2/NH2)的高效偶联。

IF 2.2 4区工程技术 Q3 PHARMACOLOGY & PHARMACY

Bioimpacts Pub Date : 2021-01-01 Epub Date: 2021-04-28 DOI:10.34172/bi.2021.34

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

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引用次数: 1

摘要

导论:磁检测技术的进一步发展可能在提高乙型肝炎病毒(HBV)感染检测和定量的敏感性方面具有重要意义。本研究基于(3-氨基丙基)三乙氧基硅烷(APTES)修饰的核壳磁性纳米颗粒(NPs)(命名为Fe3O4@SiO2/NH2)和壳聚糖修饰的磁性NPs (Fe3O4@CS)，制备和表征了一种新的生物纳米复合材料。方法:第一步分别用二氧化硅、APTES (Fe3O4@SiO2/NH2)和壳聚糖(Fe3O4@CS)对Fe3O4进行改性。山羊抗hbsag抗体被两种不同的方案激活:高碘酸钠和EDC-NHS。然后将所得复合物与活化的山羊抗hbsag IgG偶联。用外置磁铁收集生物超磁NPs (BSMNPs)，用Bradford法分析剩余溶液，检测BSMNPs表面附着抗体的量。结果:高碘酸钠法可激活抗体15 ~ 17µg，固定在1mg超磁纳米颗粒(SMNPs)上。然而，在EDC-NHS方法中，8-10µg抗体与1 mg SMNPs偶联。利用酶联免疫吸附试验(ELISA)将所得的生物磁性NPs与HBsAg靶标相互作用。约1µg抗原附着在1 mg SMNPs上，表明制备的材料适用于HBsAg的检测范围。结论:在本研究中，我们利用一种有效的偶联策略，开发了新的抗体偶联磁性NPs用于检测HBsAg。结果表明，Fe3O4@SiO2/NH2的结合能力与市售产品相当。我们设计的将抗hbsag抗体偶联到磁性纳米粒子的方法为生产高容量的磁性纳米粒子开辟了道路。

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

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

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.

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来源期刊

Bioimpacts Pharmacology, Toxicology and Pharmaceutics-Pharmaceutical Science

CiteScore

4.80

自引率

7.70%

发文量

审稿时长

5 weeks

期刊介绍： BioImpacts (BI) is a peer-reviewed multidisciplinary international journal, covering original research articles, reviews, commentaries, hypotheses, methodologies, and visions/reflections dealing with all aspects of biological and biomedical researches at molecular, cellular, functional and translational dimensions.