Development of Nanobodies Targeting Peste des Petits Ruminants Virus: The Prospect in Disease Diagnosis and Therapy
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 serologic...
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MDPI AG
2021-07-01
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| Series: | Animals |
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| Online Access: | https://www.mdpi.com/2076-2615/11/8/2206 |
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| author | Edson Kinimi Serge Muyldermans Cécile Vincke Steven Odongo Richard Kock Satya Parida Mana Mahapatra Gerald Misinzo |
| author_facet | Edson Kinimi Serge Muyldermans Cécile Vincke Steven Odongo Richard Kock Satya Parida Mana Mahapatra Gerald Misinzo |
| author_sort | Edson Kinimi |
| collection | DOAJ |
| description | 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., <i>Camelidae</i>, <i>Suidae</i>, and <i>Bovinae</i>), 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 (<i>Vicugna pacos</i>) 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 <i>ccdB</i> 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. |
| first_indexed | 2024-03-10T09:05:46Z |
| format | Article |
| id | doaj.art-eada10432c924d91ae56a6c8c4b6492d |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-10T09:05:46Z |
| publishDate | 2021-07-01 |
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| record_format | Article |
| series | Animals |
| spelling | doaj.art-eada10432c924d91ae56a6c8c4b6492d2023-11-22T06:28:45ZengMDPI AGAnimals2076-26152021-07-01118220610.3390/ani11082206Development of Nanobodies Targeting Peste des Petits Ruminants Virus: The Prospect in Disease Diagnosis and TherapyEdson Kinimi0Serge Muyldermans1Cécile Vincke2Steven Odongo3Richard Kock4Satya Parida5Mana Mahapatra6Gerald Misinzo7SACIDS Africa Centre of Excellence for Infectious Diseases, SACIDS Foundation for One Health, Sokoine University of Agriculture, P.O. Box 3297, Morogoro 25523, TanzaniaLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, BelgiumLaboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, BelgiumDepartment of Biotechnical and Diagnostic Sciences, College of Veterinary Medicine, Animal Resources and Biosecurity (COVAB), Makerere University, Kampala 7062, UgandaThe Royal Veterinary College, University of London, Hawkshead Lane, North Mymms, Hatfield AL9 7TA, UKSACIDS Africa Centre of Excellence for Infectious Diseases, SACIDS Foundation for One Health, Sokoine University of Agriculture, P.O. Box 3297, Morogoro 25523, TanzaniaThe Pirbright Institute, Ash Road, Pirbright, Woking GU24 0NF, UKSACIDS Africa Centre of Excellence for Infectious Diseases, SACIDS Foundation for One Health, Sokoine University of Agriculture, P.O. Box 3297, Morogoro 25523, TanzaniaPeste 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., <i>Camelidae</i>, <i>Suidae</i>, and <i>Bovinae</i>), 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 (<i>Vicugna pacos</i>) 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 <i>ccdB</i> 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.https://www.mdpi.com/2076-2615/11/8/2206peste des petits ruminants viruscamelidalpacananobodyheavy-chain only antibodydiagnosis |
| spellingShingle | Edson Kinimi Serge Muyldermans Cécile Vincke Steven Odongo Richard Kock Satya Parida Mana Mahapatra Gerald Misinzo Development of Nanobodies Targeting Peste des Petits Ruminants Virus: The Prospect in Disease Diagnosis and Therapy Animals peste des petits ruminants virus camelid alpaca nanobody heavy-chain only antibody diagnosis |
| title | Development of Nanobodies Targeting Peste des Petits Ruminants Virus: The Prospect in Disease Diagnosis and Therapy |
| title_full | Development of Nanobodies Targeting Peste des Petits Ruminants Virus: The Prospect in Disease Diagnosis and Therapy |
| title_fullStr | Development of Nanobodies Targeting Peste des Petits Ruminants Virus: The Prospect in Disease Diagnosis and Therapy |
| title_full_unstemmed | Development of Nanobodies Targeting Peste des Petits Ruminants Virus: The Prospect in Disease Diagnosis and Therapy |
| title_short | Development of Nanobodies Targeting Peste des Petits Ruminants Virus: The Prospect in Disease Diagnosis and Therapy |
| title_sort | development of nanobodies targeting peste des petits ruminants virus the prospect in disease diagnosis and therapy |
| topic | peste des petits ruminants virus camelid alpaca nanobody heavy-chain only antibody diagnosis |
| url | https://www.mdpi.com/2076-2615/11/8/2206 |
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