Development and Utilization of VHH Antibodies Derived from Camelus Dromedarius Against Foot-and-Mouth Disease Virus

被引:8
作者
Dash, Lipsa [1 ]
Subramaniam, Saravanan [1 ]
Khulape, Sagar A. [1 ]
Prusty, Bikash Ranjan [1 ]
Pargai, Kamal [1 ]
Narnaware, Shirish D. [2 ]
Patil, Niteen V. [2 ]
Pattnaik, Bramhadev [1 ]
机构
[1] ICAR Res Complex, Directorate Foot & Mouth Dis, Naini Tal 263138, Uttarakhand, India
[2] ICAR Res Complex, Natl Res Ctr Camel, Bikaner, Rajasthan, India
关键词
ELISA; FMDV; heavy-chain antibodies; immune library; 2XYT: Amp-Glu medium; CDR: complementary determining regions; DAS-ELISA: double-antibody sandwitch ELISA; FR1: framework region 1; FR2: framework region 2; LB agar: Luria Bertani agar; OPD: Orthophenylene-diamine dihydrochloride; PBS: phosphate-buffered saline; PBST: phosphate-buffered saline with Tween20; SMP: skimmed milk powder; TRX: Thioredoxin tag; NANOBODIES;
D O I
10.1080/10495398.2018.1433191
中图分类号
S8 [畜牧、 动物医学、狩猎、蚕、蜂];
学科分类号
0905 ;
摘要
Foot-and-mouth disease (FMD) is an acute, highly contagious, and economically devastating viral disease of domestic and wildlife species. For effective implementation of FMD control program, there is an imperative need for developing a rapid, sensitive, and specific diagnostics which help in the identification of serotypes involved in the outbreaks. The humoral immune response of the Camelidae is unique since in these animals 75% of circulating antibodies are constituted by heavy-chain antibodies and 25% are conventional immunoglobulin with two identical heavy chains. In the present study, we developed and characterized FMD virus-specific single-domain heavy-chain antibodies (VHHs) against inactivated whole-virus antigens of FMDV serotypes O (INDR2/1975), A (IND40/2000), and Asia 1 (IND63/1972) vaccine strains. After six rounds of panning and enrichment, these VHHs were stably expressed in Escherichia coli cells. The VHHs directed against outer capsid proteins of FMD virus were successfully utilized as the capture antibody in liquid-phase blocking ELISA (LPBE) thus replacing rabbit coating antibodies. Our study demonstrated the utility of FMD virus-specific VHHs as potential candidates in FMD research and diagnostic application.
引用
收藏
页码:57 / 62
页数:6
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