Construction of a camelid VHH yeast two-hybrid library and the selection of VHH against haemagglutinin-neuraminidase protein of the Newcastle disease virus
| Autor: | Sa Xiao, Enqi Du, Dandan Liu, Xiaolong Gao, Xiangyun Hu, Xinglong Wang, Haixin Wang, Xudong Chang, Ruyi Dang, Zengqi Yang, Lina Tong |
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| Jazyk: | angličtina |
| Předmět: |
0301 basic medicine
Camelus animal structures Hemagglutination viruses 030106 microbiology Molecular Sequence Data Immunoglobulin Variable Region Newcastle disease virus VHH Newcastle disease Virus Neutralization Epitope 03 medical and health sciences Two-Hybrid System Techniques Animals HN Protein Amino Acid Sequence Gene Library General Veterinary biology Immunogenicity Yeast two-hybrid General Medicine biology.organism_classification Virology veterinary(all) 030104 developmental biology biology.protein Female Antibody Immunoglobulin Heavy Chains Research Article |
| Zdroj: | BMC Veterinary Research |
| ISSN: | 1746-6148 |
| DOI: | 10.1186/s12917-016-0664-1 |
| Popis: | Background Newcastle disease (ND), which is caused by the Newcastle disease virus (NDV), is one of the most important avian diseases in poultry. Since its discovery in 1926, ND has caused great economic losses to the world poultry industry and remains a threat to chickens and wild birds. Although a stringent vaccination policy is widely adopted to control ND, ND outbreaks still occur, and virulent NDV is sporadically isolated from chickens and wild birds. To study the pathogenesis of ND and provide tools to prevent its prevalence, novel antibody fragments should be developed. The variable domains of the heavy chain of the heavy-chain antibodies (VHH) are the smallest naturally occurring antibodies derived from camelid heavy-chain antibodies. The comparatively small size, high affinity, high solubility, low immunogenicity and ability to bind epitopes inaccessible to conventional antibodies of VHH make them ideal candidates for a considerable number of therapeutic and biotechnological applications. However, an anti-NDV VHH has not been reported to date. Results In this study, a VHH yeast two-hybrid library was constructed from NDV vaccine immunized C. bactrianus, and seven VHH fragments to the haemagglutinin-neuraminidase (HN) protein of NDV were successfully screened and characterized for the first time. These selected VHH clones were all expressed as soluble protein in E. coli. ELISA, dot blot, immunocytochemistry and pull down results showed that the screened VHHs could interact with NDV virion, among which five had neutralizing activity. In addition, the seven VHHs could inhibit the haemagglutination activity of different NDV strains. Conclusions We constructed an NDV-immunized VHH yeast two-hybrid library and screened and characterized seven VHHs targeting NDV HN protein for the first time. The seven VHHs may have great potential for NDV diagnosis, pathogenesis and therapeutics. Electronic supplementary material The online version of this article (doi:10.1186/s12917-016-0664-1) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
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