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Abstract
Purpose
COVID-19 is rampant throughout the world, which has caused great damage to human lives and seriously hindered the development of the global economy. Aiming at the treatment of SARS-CoV-2, in this study, we proposed a novel fenobody strategy based on ferritin (Fe) self-assembly technology.
Methods
The neutralizing nanobody H11-D4 of SARS-CoV-2 fused to the C-terminus of end-modified human ferritin was expressed in E. coli and silkworm baculovirus expression systems. A large number of nanoparticles were successfully self-assembled in silkworms, while relatively few nanoparticles can be observed in the treated products from E. coli by electron microscopy. Subsequently, the fenobody’s expression level and neutralizing activity were then evaluated.
Results
The results showed that the IC50 of H11-D4 and fenobody Fe-H11-D4 expressed in E. coli were 171.1 nmol L−1 and 20.87 nmol L−1, respectively. However, the IC50 of Fe-HD11-D4 expressed in silkworms was 1.46 nmol L−1 showing better neutralization activity.
Conclusion
Therefore, fenobodies can be well self-assembled in silkworm baculovirus expression system, and ferritin self-assembly technology can effectively improve nanobody neutralization activity.
Acknowledgments
This work was supported by the National Key Research and Development Program of China (No. 2022YFD1300700), the National Natural Sciences Foundation of China (No. 32372952, 32072796, 32002236), the Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences (CAAS-ZDRW202305), and the Regional Demonstration Project of Huzhou Academy of Agricultural Sciences Alliance (No. 2022SJLM10).
Disclosure
National Invention Patents, SARS-CoV-2 neutralizing nanobody, self-assembled ferritin fusion nanobody and preparation method and application, Li Yinü, Zhang Zhifang, Liu Xingjian, Gao Xintao, Yi Yongzhu, Wang Haining, 20220614, CN1, 20220902, ZL 2022 1 0663315.9. The authors report no other conflicts of interest in this work.