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.
| Original language | English |
|---|---|
| Article number | 2206 |
| Number of pages <span style="color:red"p> <font size="1.5"> ✽ </span> </font> | 13 |
| Journal | Animals |
| Volume | 11 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - Aug 2021 |
Bibliographical note
Funding Information:Funding: E.K. is recipient of a scholarship from the Government of the United Republic of Tanzania through the World Bank (WB-ACE II Grant PAD1436, IDA credit 5799-TZ) to the SACIDS-Centre of Excellence for Infectious Diseases of Humans and Animals in Eastern and Southern Africa (SACIDS-ACE) at the SACIDS Foundation for One Health of the Sokoine University of Agriculture, Tanzania, East Africa. The nanobody Ablynx technology and equipment were generously provided by the Laboratory of Cellular and Molecular immunology at Vrije Universiteit Brussel, Brussels, Belgium. The cost of PPRV antigens for biopanning and article processing charges were funded by The Pirbright Institute through the Biotechnology and Biological Sciences Research Council (BBSRC), grant numbers BB/L013657/1 and BB/T004096/1.
Funding Information:
Acknowledgments: The authors wish to thank the Government of the United Republic of Tanzania and World Bank for providing a scholarship grant to E.K. We also extend our sincere thanks to Ema Rom\u00E3o for her guidance at some points during the library construction, as well as Maxine Crauwels, Christopher Kariuki, Els Lebegge, Sam Massa, Francisco J. Morales-Yanez, Joar Pinto Torres, Francis Saetens, Nadia Abou and Carol Kagia for their technical assistance and logistic support at Vrije Universiteit Brussel, Brussels, Belgium. We are indebted to Paul Gwakisa for his excellent discussions during conceptualization and operationalization of nanobody research. Equally, we express our deep and sincere gratitude to Michael Baron for his tireless and invaluable guidance during outsourcing of PPRV antigens. Our special thanks to the Executive Director of the SACIDS Foundation for One Health, Mark M. Rweyemamu, for his strategic guidance and immense contribution towards infectious diseases of public health and veterinary importance.
Funding Information:
E.K. is recipient of a scholarship from the Government of the United Republic of Tanzania through the World Bank (WB-ACE II Grant PAD1436, IDA credit 5799-TZ) to the SACIDS-Centre of Excellence for Infectious Diseases of Humans and Animals in Eastern and Southern Africa (SACIDS-ACE) at the SACIDS Foundation for One Health of the Sokoine University of Agriculture, Tanzania, East Africa. The nanobody Ablynx technology and equipment were generously provided by the Laboratory of Cellular and Molecular immunology at Vrije Universiteit Brussel, Brussels, Belgium. The cost of PPRV antigens for biopanning and article processing charges were funded by The Pirbright Institute through the Biotechnology and Biological Sciences Research Council (BBSRC), grant numbers BB/L013657/1 and BB/T004096/1.
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.