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Generation and characterisation of recombinant FMDV antibodies: Applications for advancing diagnostic and laboratory assays

DOI: 10.1371/journal.pone.0201853 DOI Help

Authors: Gareth Shimmon (The Pirbright Institute) , Abhay Kotecha (University of Oxford) , Jingshan Ren (University of Oxford) , Amin S. Asfor (The Pirbright Institute) , Joseph Newman (The Pirbright Institute) , Stephen Berryman (The Pirbright Institute) , Eleanor M. Cottam (The Pirbright Institute) , Sarah Gold (The Pirbright Institute) , Toby J. Tuthill (The Pirbright Institute) , Donald P. King (The Pirbright Institute) , Emiliana Brocchi (Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna) , Andrew M. Q. King (The Pirbright Institute) , Ray Owens (University of Oxford) , Elizabeth E. Fry (University of Oxford) , David I. Stuart (University of Oxford; Diamond Light Source) , Alison Burman (The Pirbright Institute) , Terry Jackson (The Pirbright Institute)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Plos One , VOL 13

State: Published (Approved)
Published: August 2018

Open Access Open Access

Abstract: Foot-and-mouth disease (FMD) affects economically important livestock and is one of the most contagious viral diseases. The most commonly used FMD diagnostic assay is a sandwich ELISA. However, the main disadvantage of this ELISA is that it requires anti-FMD virus (FMDV) serotype-specific antibodies raised in small animals. This problem can be, in part, overcome by using anti-FMDV monoclonal antibodies (MAbs) as detecting reagents. However, the long-term use of MAbs may be problematic and they may need to be replaced. Here we have constructed chimeric antibodies (mouse/rabbit D9) and Fabs (fragment antigen-binding) (mouse/cattle D9) using the Fv (fragment variable) regions of a mouse MAb, D9 (MAb D9), which recognises type O FMDV. The mouse/rabbit D9 chimeric antibody retained the FMDV serotype-specificity of MAb D9 and performed well in a FMDV detection ELISA as well as in routine laboratory assays. Cryo-electron microscopy analysis confirmed engagement with antigenic site 1 and peptide competition studies identified the aspartic acid at residue VP1 147 as a novel component of the D9 epitope. This chimeric expression approach is a simple but effective way to preserve valuable FMDV antibodies, and has the potential for unlimited generation of antibodies and antibody fragments in recombinant systems with the concomitant positive impacts on the 3Rs (Replacement, Reduction and Refinement) principles.

Journal Keywords: Enzyme-linked immunoassays; Antibodies; Foot and mouth disease; Viral packaging; Elution; Integrins; Viral structure; Electron cryo-microscopy

Subject Areas: Biology and Bio-materials


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