Key result
Engineered recombinant scFv-mAb successfully detects FMDV serotype O via indirect sandwich ELISA.
The engineered recombinant scFv-mAb expressed in E. coli successfully detected serotype O of the Foot-and-mouth disease virus, demonstrating potential for diagnostic applications.
May support FMDV diagnostic development; leaves open validation before veterinary adoption.
Introduction: Foot-and-mouth disease (FMD) is a highly contagious viral disease caused by the FMD virus (FMDV), which belongs to the Aphthovirus genus in the Picornaviridae family. FMDV is a highly variable RNA virus, and there is limited cross-protection between serotypes and strains. The disease can have devastating economic, social, and environmental impacts. The FMDV capsid is composed of VP1-VP4 structural proteins, with VP1 being the most abundant protein consisting of 213 amino acids. The G-H loop (residues 141-160) of the VP1 capsid protein is highly variable and serves as the main antigenic region. It contains a conserved triplet of amino acids, Arg-Gly-Asp (RGD), which induces the production of protective antibodies against various FMDV types. Monoclonal antibodies (mAbs) play a crucial role in detecting and serotyping FMDV in pathological samples, as well as evaluating protection post-vaccination against FMD.Materials & Methods: This study explores the expression and function of an engineered recombinant single-chain variable fragment (scFv-mAb) in Escherichia coli BL21 (DE3) Rosetta strain as a cost-effective prokaryotic system with high yield. The scFv-mAb gene was inserted into the pET28a (+) expression plasmid. E. coli cells were transformed with the plasmid, induced with 0.5 mM IPTG, and incubated at 37 ˚C for 12 hours. The protein was purified using a Ni2+-NTA resin column and analyzed by 12% SDS-PAGE for quality assessment. The efficiency and functionality of the scFv-mAb were confirmed using an indirect sandwich (capture) enzyme-linked immunosorbent assay (ELISA).Results: The purified scFv-mAb concentration was determined to be approximately 2.00 mg/mL using the Bradford assay under optimal conditions. Each well was coated with 400 ng of scFv-mAb based on checkerboard results and the mean of negative serum at a 1:10 dilution. The indirect sandwich ELISA assay yielded an optical density (OD) signal range of 0.3 to 1.5 at a 450 nm wavelength in different positive control treatments.Conclusion: The ELISA results showed that the scFv-mAb fragment successfully detected serotype O of FMDV. Further research could confirm the potential of this recombinant antibody for broader commercial applications in the future.
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Helalinasab et al. (2026) studied Foot-and-mouth disease (FMD) virus detection. Recombinant scFv-mAb against VP1 capsid protein vs. Negative control serum was evaluated on Detection of FMDV serotype O via indirect sandwich ELISA (OD at 450 nm). The engineered recombinant scFv-mAb successfully detected FMDV serotype O in an indirect sandwich ELISA, yielding an optical density signal range of 0.3 to 1.5 at 450 nm.
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