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Tytuł:
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Selection and identification of single-domain antibody against Peste des Petits Ruminants virus.
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Autorzy:
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Liu D; Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
Li L; Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
Cao X; Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
Wu J; Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China. .
Du G; Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
Shang Y; Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.
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Źródło:
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Journal of veterinary science [J Vet Sci] 2021 Jul; Vol. 22 (4), pp. e45. Date of Electronic Publication: 2021 Jun 02.
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Typ publikacji:
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Journal Article
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Język:
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English
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Imprint Name(s):
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Original Publication: Seoul : Korean Society of Veterinary Science, 2000-
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MeSH Terms:
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Antibody Specificity*
Camelus*
Viral Vaccines*
Antibodies, Viral/*immunology
Peste-des-Petits-Ruminants/*prevention & control
Peste-des-petits-ruminants virus/*immunology
Animals ; Antibody Affinity ; Chlorocebus aethiops ; Cloning, Molecular ; Female ; Gene Library ; RNA, Messenger ; Vero Cells
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References:
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Grant Information:
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China Research and demonstration on rapid diagnosis technology of local high incidence; China China Agriculture Research System of MOF and MARA
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Contributed Indexing:
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Keywords: Peste des petits ruminants virus; cDNA library; single-domain antibody
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Substance Nomenclature:
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0 (Antibodies, Viral)
0 (RNA, Messenger)
0 (Viral Vaccines)
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Entry Date(s):
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Date Created: 20210625 Date Completed: 20211006 Latest Revision: 20211006
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Update Code:
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20240105
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PubMed Central ID:
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PMC8318796
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DOI:
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10.4142/jvs.2021.22.e45
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PMID:
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34170088
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Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Single-domain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility.
Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV.
Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2.
Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 10⁶ cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein.
Conclusions: The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.
Competing Interests: The authors declare no conflicts of interest.
(© 2021 The Korean Society of Veterinary Science.)
