Viruses (Feb 2025)

Neutralizing Antibody Screening Using NanoBiT-Based Virus-like Particles of Foot-and-Mouth Disease Type Asia1 Enhances Biosafety and Sensitivity

  • Hyejin Kim,
  • Dong-Wan Kim,
  • Giyoun Cho,
  • Ji-Hyeon Hwang,
  • Yeonrae Chae,
  • Taejun Kim,
  • Jae Young Kim,
  • Young-Joon Ko,
  • Jong-Hyeon Park,
  • Yoon-Hee Lee,
  • Sung-Han Park

DOI
https://doi.org/10.3390/v17030337
Journal volume & issue
Vol. 17, no. 3
p. 337

Abstract

Read online

Background/Objectives: Foot-and-mouth disease (FMD) is a highly contagious class 1 animal disease that affects cloven-hoofed animals, such as cattle, pigs, and goats. Diagnosis and research on live FMD virus (FMDV) typically require biosafety level 3 facilities, which are challenging to maintain due to strict protocols and high costs. The development of NanoBiT-based assays has accelerated in response to the coronavirus disease pandemic, providing safer alternatives for viral research, and is now applicable for general laboratories. This study aimed to develop a NanoBiT-based virus-like particle (VLP) assay for the rapid and safe screening of neutralizing antibodies against FMDV Asia1 Shamir (AS). Methods: We developed an AS VLP with an inserted HiBiT tag that enabled the detection of entry into LgBiT cells through luminescence signals. Results: HiBiT-tagged AS VLPs mixed with anti-serum and introduced into LgBiT-expressing cells led to a reduction in luciferase activity. Therefore, we established a NanoBiT-based viral neutralizing antibody test (VNT) that demonstrated a high correlation (R2 = 0.881) with the traditional gold standard VNT. Conclusions: The assay demonstrated high sensitivity and could be performed in BL-2 facilities, offering a safer and more efficient alternative to traditional assays while reducing the need to handle live viruses in high-containment facilities. This method provides a valuable tool for rapid screening of neutralizing antibodies and can be adapted for broader applications in FMDV research.

Keywords