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.
Highlights
Foot-and-mouth disease virus (FMDV) causes significant foot-and-mouth disease for cloven-hoofed animals [1]
When excluding the cross-reactive type Asia1 serum, these results indicate BvO17 and BvA3 can be used for serotype-specific detection of FMDV antibodies
We successfully isolated bovine anti-FMDV antibodies by phage display using scFv library generated from peripheral blood lymphocytes (PBL) of cattle in a local slaughterhouse
Summary
Foot-and-mouth disease virus (FMDV) causes significant foot-and-mouth disease for cloven-hoofed animals [1]. When the PI value is greater than 50%, the serum turns out to be FMDV type O antibody positive Kit performance such as sensitivity and specificity is very dependent on the choice of MAb. The concern about using MAb reagent in cELISA is how well the antibody recognizing a single epitope can represent overall antibody response against diverse epitopes on FMDV capsid. Extensive studies on antigenic features of FMDV, especially type O, have identified major neutralizing epitopes recognized by the mouse MAbs [11,12,13] Even though these epitope regions are suggested to be recognized by bovine antibodies [14,15], the relative preference of each epitope region and fine epitope structure may be different in cattle.
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