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Fe3O4@SiO2-Protein A-oHSV/CD63 Ab for Capturing Virus and Exosomes

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Abstract

Protein A modified magnetic spheres (Fe3O4@SiO2-Protein A) with 22.7 emu/g saturation magnetization were prepared and characterized. These spheres had a diameter of 10 nm and was stable up to 229.2 °C, and can bond 19.694 mg/g of Protein A. The Fe3O4@SiO2-Protein A was firstly combined with oHSV-BJ-2-A antibody to capture oHSV from disease venom, and the amount of binding virus was 1.92 × 107 to 3.44 × 107 PFU/g. Fe3O4@SiO2-Protein A was modified with CD63 antibody, which was further fixed using dissuccinimide octylate (DSS) as a crosslinking agent. The prepared Fe3O4@SiO2-Protein A-CD63 Ab was useful in isolating and enriching exosomes from cell supernatant. Compared with commercial kits, Fe3O4@SiO2-Protein A-CD63 Ab demonstrated a better purification effect, which was successfully monitored using capillary electrophoresis. The total content of exosomes protein extracted using Fe3O4@SiO2-Protein A-CD63 was 3.4 mg/g, which was obviously higher than that reported in some studies. In addition, Fe3O4@SiO2-Protein A-CD63 Ab also showed its repeatability through bonding and elution of 5 cycles, which was effective in cost saving. These indicated Fe3O4@SiO2-Protein A-CD63 Ab had the potential for large-scale purification of exosomes in practical applications.

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Acknowledgements

Authors gratefully acknowledge the funding supports from National Natural Science Foundation of China (21605112).

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Author notes

  1. Wenqing Zheng, Pingyi Zheng and Ran Zhao have contributed equally to this work.

Authors and Affiliations

  1. Tianjin Key Laboratory for Modern Drug Delivery and High-Efficiency, Collaborative Innovation Center of Chemical Science and Engineering, School of Pharmaceutical Science and Technology, Faculty of Medicine, Tianjin University, Tianjin, 300072, China

    Wenqing Zheng, Pingyi Zheng, Ran Zhao, Xinyu Xu, Xiao Zhang, Xiaoqian Yuan, Ying Xu, Zichuan Liu & Youxin Li

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  1. Wenqing Zheng
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  2. Pingyi Zheng
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  3. Ran Zhao
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  4. Xinyu Xu
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  5. Xiao Zhang
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  6. Xiaoqian Yuan
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  7. Ying Xu
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  8. Zichuan Liu
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Contributions

Wenqing Zheng and Pingyi Zheng conceived and designed the experiments; Wenqing Zheng, Pingyi Zheng, and Ran Zhao performed the experiments and analyzed the data; Xinyu Xu, Xiao Zhang, Xiaoqian Yuan helped to revise the paper; Ying Xu cultured cells; Youxin Li provided the concept of this research and managed all the experimental and writing process as the corresponding authors; Zichuan Liu guided the cell culture and revised the paper. All authors discussed the results, commented the paper and agreed to the published version of the paper.

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Correspondence to Zichuan Liu or Youxin Li.

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Zheng, W., Zheng, P., Zhao, R. et al. Fe3O4@SiO2-Protein A-oHSV/CD63 Ab for Capturing Virus and Exosomes. J. Anal. Test. (2024). https://doi.org/10.1007/s41664-024-00310-5

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