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Background <p>Foot-and-mouth disease (FMD), caused by foot-and-mouth disease virus (FMDV), is an acute, highly contagious, and economically devastating disease affecting cloven-hoofed animals worldwide. Vaccination remains the cornerstone of FMD control strategies. To increase the production yield and structural stability of virus-like particles (VLPs), we investigated the effects of the N-terminal truncation of VP0 on the soluble co-expression of three capsid proteins, and the stability of the assembled particles.</p> Results <p>A series of VP0 variants with progressive N-terminal truncations (5–20 amino acids) were co-expressed with VP3 and VP1 via a single plasmid in the <i>Escherichia coli</i> (<i>E. coli</i>) expression system by fusing these proteins with a small ubiquitin-like modifier (SUMO) tag. The assembly characteristics of the co-expressed fusion proteins were systematically characterized. Our findings show that all engineered fusion proteins were successfully co-expressed, and truncations between residues 10 and 20 yielded significantly higher expression levels and purification efficiencies than those of full-length VP0. The 031-VP0 dN10, 031-VP0 dN15, and 031-VP0 dN20 exhibited increased stability of the assembly and retained the native morphology. An in vitro binding assay for neutralizing antibodies against FMDV indicated that all VLPs maintained wild-type-like antigenic properties, demonstrating preserved conformational epitopes. Pigs immunized with VLPs-based vaccines exhibited humoral and cellular immune responses, with the 031-VP0 dN20 variant eliciting significantly stronger cellular immunity.</p> Conclusions <p>Collectively, our findings establish a robust platform for the efficient production of structurally stable FMDV VLPs in <i>E. coli</i>, providing a valuable framework for developing soluble expression systems for other FMDV serotypes.</p>

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N-terminal truncation of the foot-and-mouth disease virus VP0 protein promotes soluble co-expression and particle assembly stability of viral capsid proteins in Escherichia coli

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年10月10日

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  • Suling Zhang,
  • Siyu Wu,
  • Yan Liu,
  • Xiaolin Geng,
  • Yuxin Huang,
  • Yanan Yue,
  • He Yan,
  • Peng Wu,
  • Yurui Wang,
  • Kegong Tian,
  • Aihua Wang,
  • Wenqiang Pang

摘要

Background

Foot-and-mouth disease (FMD), caused by foot-and-mouth disease virus (FMDV), is an acute, highly contagious, and economically devastating disease affecting cloven-hoofed animals worldwide. Vaccination remains the cornerstone of FMD control strategies. To increase the production yield and structural stability of virus-like particles (VLPs), we investigated the effects of the N-terminal truncation of VP0 on the soluble co-expression of three capsid proteins, and the stability of the assembled particles.

Results

A series of VP0 variants with progressive N-terminal truncations (5–20 amino acids) were co-expressed with VP3 and VP1 via a single plasmid in the Escherichia coli (E. coli) expression system by fusing these proteins with a small ubiquitin-like modifier (SUMO) tag. The assembly characteristics of the co-expressed fusion proteins were systematically characterized. Our findings show that all engineered fusion proteins were successfully co-expressed, and truncations between residues 10 and 20 yielded significantly higher expression levels and purification efficiencies than those of full-length VP0. The 031-VP0 dN10, 031-VP0 dN15, and 031-VP0 dN20 exhibited increased stability of the assembly and retained the native morphology. An in vitro binding assay for neutralizing antibodies against FMDV indicated that all VLPs maintained wild-type-like antigenic properties, demonstrating preserved conformational epitopes. Pigs immunized with VLPs-based vaccines exhibited humoral and cellular immune responses, with the 031-VP0 dN20 variant eliciting significantly stronger cellular immunity.

Conclusions

Collectively, our findings establish a robust platform for the efficient production of structurally stable FMDV VLPs in E. coli, providing a valuable framework for developing soluble expression systems for other FMDV serotypes.