Phage Display Screening of Bovine Antibodies to Foot-and-Mouth Disease Virus and Their Application in a Competitive ELISA for Serodiagnosis

Abstract

For serodiagnosis of foot-and-mouth disease virus (FMDV), monoclonal antibody (MAb)-based competitive ELISA (cELISA) is commonly used since it allows simple and reproducible detection of antibody response to FMDV. However, the use of mouse-origin MAb as a detection reagent is questionable, as antibody responses to FMDV in mice may differ in epitope structure and preference from those in natural hosts such as cattle and pigs. To take advantage of natural host-derived antibodies, a phage-displayed scFv library was constructed from FMDV-immune cattle and subjected to two separate pannings against inactivated FMDV type O and A. Subsequent ELISA screening revealed high-affinity scFv antibodies specific to a serotype (O or A) as well as those with pan-serotype specificity. When BvO17, an scFv antibody specific to FMDV type O, was tested as a detection reagent in cELISA, it successfully detected FMDV type O antibodies for both serum samples from vaccinated cattle and virus-challenged pigs with even higher sensitivity than a mouse MAb-based commercial FMDV type O antibody detection kit. These results demonstrate the feasibility of using natural host-derived antibodies such as bovine scFv instead of mouse MAb in cELISA for serological detection of antibody response to FMDV in the susceptible animals.

Principles of antibodies with ultralong complementarity-determining regions and picobodies

“…In contrast to yeast and mammalian surface display, phage display has had less success in the discovery of ulCDR-H3s. Three studies employing phage display to isolate antigen-specific bovine antibodies yielded only binders with conventional CDR-H3s [39][40][41] .…”

The Biotechnological Application of Bacteriophages: What to Do and Where to Go in the Middle of the Post-Antibiotic Era

“…This adaptable tool fuels innovative strategies for disease detection and the exploration of biomarkers. Once engineered, these modified phages are useful in diverse diagnostic applications, offering sensitive and precise detection of disease-related entities in clinical samples, ranging from minute targets such as viruses to substantial entities such as cancer cells ( Table 1 ) [ 182 , 183 , 184 , 185 , 186 ].…”

Screening of sperm antigen epitopes by phage display technique and its preliminary clinical application

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