Abstract
Foot-and-mouth disease (FMD), caused by FMD virus (FMDV), is a highly contagious viral disease affecting cloven-hoofed animals. Camelids have a unique immunoglobulin profile, with the smallest functional heavy-chain antibodies (sdAb or VHH) naturally devoid of light chains with antigen-binding capacity. We screened and characterized five sdAbs against FMDV by immunized library from C. bactrianus with Asia 1 virus-like particles (VLPs). Three of five recombinant sdAbs were stably expressed in E.coli, remained highly soluble, and were serotype-specific for VP1 protein of FMDV Asia 1 by ELISA. These failed to completely neutralize the Asia 1 virus. According to the KD value of binding affinity to three sdAbs, which ranged from 0.44 to 0.71 nm by SPR, sdAb-C6 was selected and conjugated with Zn/CdSe quantum dots (QDs) to form a QDs-C6 probe, which was used to trace and image the subcellular location of FMDV in BHK-21 cells. The results show that FMD virions were observed from 3 h.p.i., and most of virions were distributed on one side of the nucleus in the cytoplasm. We demonstrate the utility of sdAbs as functionalized QDs are powerful tools for FMDV research.
Highlights
Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed livestock, caused by FMD virus (FMDV), a member of the genus Aphthovirus, in the family Picronaviridae
Detection for camels FMDV Asia 1 antibody Two Bactrian camels were appropriated from the Gobi desert before starting the experiment, and they nor their parents were vaccinated against FMDV, and were as such considered ‘‘natural specific pathogen free’’ animals
We demonstrate that mature virions can be reliably recognized and labeled with quantum dots (QDs)-sdAb probe in baby hamster kidney-21 (BHK-21) cells infected with FMDV Asia 1 up to 3 h.p.i
Summary
Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed livestock, caused by FMD virus (FMDV), a member of the genus Aphthovirus, in the family Picronaviridae. It consists of a single-stranded, positive-sense RNA genome of approximately 8.5 kb surrounded by four structural proteins (VP1 to 4), which form viral capsid proteins that are considered antigenic, in addition to 15 different, mature non-structural proteins (Nsp) [1]. In FMD-free countries, livestock, including cloven-hooved animals, are extremely susceptible to FMDV due to the non-prophylactic-vaccination policy, which results in no humoral response against FMDV. Asia has its own unique serotype, and outbreaks due to Asia 1 have been reported only sporadically in the past few decades. Analysis of Asia 1 indicates that some strains have been spread across large distances between countries in Asia within a short time [8]
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