Characterization of single-domain antibodies against Foot and Mouth Disease Virus (FMDV) serotype O from a camelid and imaging of FMDV in baby hamster kidney-21 cells with single-domain antibody-quantum dots probes
文献类型: 外文期刊
第一作者: Wang, Di
作者: Wang, Di;Yang, Shunli;Yin, Shuanghui;Shang, Youjun;Du, Ping;Guo, Jianhong;He, Jijun;Cai, Jianping;Liu, Xiangtao
作者机构:
关键词: Foot-and-mouth disease virus serotype O;Immune library;Single domain antibody;Quantum dots;Probe;Imaging
期刊名称:BMC VETERINARY RESEARCH ( 影响因子:2.741; 五年影响因子:2.955 )
ISSN: 1746-6148
年卷期: 2015 年 11 卷
页码:
收录情况: SCI
摘要: Background: Foot-and-mouth disease (FMD) is a highly contagious disease that affects cloven-hoofed animals and causes significant economic losses to husbandry worldwide. The variable domain of heavy-chain antibodies (VHHs or single domain antibodies, sdAbs) are single-domain antigen-binding fragments derived from camelid heavy-chain antibodies. Results: In this work, two sdAbs against FMD virus (FMDV) serotype O were selected from a camelid phage display immune library and expressed in Escherichia coli. The serotype specificity and affinity of the sdAbs were identified through enzyme-linked immunosorbent assay and surface plasmon resonance assay. Moreover, the sdAbs were conjugated with quantum dots to constitute probes for imaging FMD virions. Results demonstrated that the two sdAbs were specific for serotype O and shared no cross-reactivity with serotypes A and Asia 1. The equilibrium dissociation constant (KD) values of the two sdAbs ranged from 6.23 nM to 8.24 nM, which indicated high affinity to FMDV antigens. Co-localization with the sdAb-AF488 and sdAb-QD probes indicated the same location of FMDV virions in baby hamster kidney-21 (BHK-21) cells. Conclusions: sdAb-QD probes are powerful tools to detect and image FMDV in BHK-21 cells.
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