Bovine single chain Fv antibody inhibits bovine herpesvirus-1 infectivity by targeting viral glycoprotein D
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Glycoprotein D (gD) of bovine herpesvirus-1 (BoHV-1) is essential for attachment and penetration of cells during infection and is a major target for neutralizing antibodies during an adaptive immune response. Currently there are no recombinant antibodies capable of binding gD epitopes for use in treating BoHV-1 infection. In this study, a bovine scFv gene derived from a hybridoma secreting monoclonal antibodies (McAbs) against the amino acid motif MEESKGYEPP of gD was expressed in E. coli. Molecular modeling, western blot and ELISA analysis showed that this scFv had a high affinity for BoHV-1 gD, with a Kd of 161.2 ± 37.58 nM and for whole BoHV-1 virus, with a Kd of 67.44 ± 16.99 nM. In addition, this scFv displayed a high affinity for BoHV-1 antigen in an ELISA and competed with BoHV-1 anti-serum in a competitive ELISA. Immunofluorescence assay (IFA) and laser confocal microscopy showed that this scFv could efficiently bind to and be internalized by BoHV-1 infected Madin-Darby bovine kidney (MDBK) cells. Importantly, this scFv was shown to inhibit BoHV-1 infectivity and to reduce the number of viral plaques by blocking viral attachment to MDBK cells. Our study suggests that this bovine single-chain antibody could be developed for use as a diagnostic and therapeutic agent against BoHV-1 infection in cattle.
KeywordsBovine herpesvirus-1 Single chain Fv antibody Glycoprotein D Neutralizing epitope Binding affinity Blocking viral attachment Virus neutralization
This work was supported by a grant from The National Key Project of Research and Development Program of China (Grant No. 2016YFD0500900), funding from Beijing Innovation Team of Technology System in Dairy Industry (award no. bjcystx-ny-3) and the Special Program on Science and Technology Innovation Capacity Building of BAAFS (award no. KJCX20170406).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All animal research was approved by the Beijing Association for Science and Technology (approval ID SYXK (Beijing) 2007-0023) and was in compliance with Beijing Laboratory Animal Welfare and Ethics guidelines as issued by the Beijing Administration Committee of Laboratory Animals. All animal studies were also performed in accordance with the Beijing Academy of Agricultural and Forestry Sciences Institutional Animal Care and Use Committee guidelines. The protocol was approved by the Committee on Experimental Animal Management of Beijing Academy of Agricultural and Forestry Sciences.
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