Development of Nanobodies Targeting Peste des Petits Ruminants Virus: The Prospect in Disease Diagnosis and Therapy

Abstract

Peste des petits ruminants virus (PPRV) causes a highly devastating disease, peste des petits ruminants (PPR) of sheep and goats, that threatens food security, small ruminant production, and the conservation of wild small ruminants in many developing countries, especially in Africa. Robust serological and molecular diagnostic tools are available to detect PPRV infection, but they were mainly developed for domestic sheep and goats. The presence of a wide host range for PPRV does present serological diagnostic challenges. New innovative diagnostic tools are needed to detect PPRV in atypical hosts (e.g., Camelidae, Suidae, and Bovinae), in wildlife ecosystems and in complex field situations. Interestingly, single-domain antigen binding fragments (nanobodies) derived from heavy-chain-only camelid antibodies have emerged as a new hope in the development of accurate, rapid, and cost-effective diagnostic tools in veterinary and biomedical fields that are suitable for low-income countries. The main objective of this study was to construct an immune nanobody library to retrieve PPRV-reactive nanobodies that enable the development of diagnostic and therapeutic nanobodies in the future. Here, a strategy was developed whereby an alpaca (Vicugna pacos) was immunized with a live attenuated vaccine strain (PPRV/N/75/1) to raise an affinity-matured immune response in the heavy-chain-only antibody classes. The nanobody gene repertoire was engineered in pMECS-GG phagemid, whereby a ccdB gene (encoding a lethal protein) was substituted by the nanobody gene. An immune nanobody library with approximately sixty-four million independent transformants was constructed, of which 100% contained an insert with the proper size of nanobody gene. Following phage display and biopanning, nine nanobodies that specifically recognise completely inactivated PPRV were identified on enzyme-linked immunosorbent assay. They showed superb potency in rapidly identifying PPRV, which is likely to open a new perspective in the diagnosis and possible treatment of PPR infection.

Complete Genome Sequencing of Field Isolates of Peste des Petits Ruminants Virus from Tanzania Revealed a High Nucleotide Identity with Lineage III PPR Viruses

“…To enable the successful eradication of PPR, rapid diagnosis remains a cornerstone for the implementation of control measures, including vaccination, quarantine and possible stamping out [36,77]. The wide host susceptibility of species to PPR presents logistic challenges around sampling, diagnosis and diagnostic protocols need to be adequately validated for atypical species affected, and the type of sample being collected [78][79][80]. The application of Oxford nanopore MinION sequencing technology demonstrated it to be an effective and rapid option for the molecular sequencing of PPRV.…”

Schistosomiasis diagnosis: Challenges and opportunities for elimination

“…Nanobodies, which are easy to produce in large scale, could be key in development of highly sensitive, specific, and affordable diagnostics, which can be used in resource-limited settings such as sub-Saharan Africa. The use of the nanobody coupled with a lateral flow device has been proposed as the probable solution to the development of cost-effective, highly sensitive, specific, and rapid immunoassays for diagnosing peste des petits ruminants [ 69 , 70 ]. The use of nanobody technology remains to be investigated as an effective in diagnosis of schistosomiasis.…”

A Novel Blocking Enzyme-Linked Immunosorbent Assay Based on a Biotinylated Nanobody for the Rapid and Sensitive Clinical Detection of Classical Swine Fever Virus Antibodies

“…Nbs have been widely used in double-antibody sandwich ELISA, competitive ELISA, and blocking ELISA (bELISA) ( 23 – 25 ). The application of Nbs can improve the specificity and sensitivity of ELISA detection, and the occurrence of a disease can be detected earlier ( 26 , 27 ). CSFV-specific Nbs have not yet been reported.…”