Wang Di, Yang Shunli, Yin Shuanghui, Shang Youjun, Du Ping, Guo Jianhong, He Jijun, Cai Jianping, Liu Xiangtao
State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu, China.
BMC Vet Res. 2015 May 22;11:120. doi: 10.1186/s12917-015-0437-2.
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.
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.
sdAb-QD probes are powerful tools to detect and image FMDV in BHK-21 cells.
口蹄疫（FMD）是一种高度传染性疾病，可感染偶蹄动物，并给全球畜牧业造成重大经济损失。重链抗体的可变区（VHHs或单域抗体，sdAbs）是源自骆驼科动物重链抗体的单域抗原结合片段。
在本研究中，从骆驼科噬菌体展示免疫文库中筛选出两种针对口蹄疫病毒（FMDV）O型的sdAbs，并在大肠杆菌中表达。通过酶联免疫吸附测定和表面等离子体共振测定确定了sdAbs的血清型特异性和亲和力。此外，将sdAbs与量子点偶联，构成用于口蹄疫病毒粒子成像的探针。结果表明，这两种sdAbs对O型具有特异性，与A型和亚洲1型无交叉反应。两种sdAbs的平衡解离常数（KD）值在6.23 nM至8.24 nM之间，表明它们与FMDV抗原具有高亲和力。sdAb-AF488和sdAb-QD探针的共定位表明口蹄疫病毒粒子在幼仓鼠肾-21（BHK-21）细胞中的位置相同。
sdAb-QD探针是检测和成像BHK-21细胞中口蹄疫病毒的有力工具。
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