Specific determination of influenza H7N2 virus based on biotinylated single-domain antibody from a phage-displayed library

doi:10.1016/j.ab.2015.09.020

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	Specific determination of influenza H7N2 virus based on biotinylated single-domain antibody from a phage-displayed library
	Gong, Xue 5; Zhu, Min 2; Li, Guanghui 2; Lu, Xiaoling 1,3,4; Wan, Yakun 2
刊名	ANALYTICAL BIOCHEMISTRY
	2016-05-01
卷号	500 页码:66-72
关键词	Influenza H7N2 virus VHH antibody Phage library Sandwich ELISA
ISSN号	0003-2697
DOI	10.1016/j.ab.2015.09.020
文献子类	Article
英文摘要	The unpredicted spread of avian influenza virus subtype H7N2 in the world is threatening animals and humans. Specific and effective diagnosis and supervision are required to control the influenza. However, the existing detecting methods are laborious, are time-consuming, and require appropriate laboratory facilities. To tackle this problem, we isolated VHH antibodies against the H7N2 avian influenza virus (AIV) and performed an enzyme-linked immunosorbent assay (ELISA) to detect the H7N2 virus. To obtain VHH antibodies with high affinity and specificity, a camel was immunized. A VHH antibody library was constructed in a phage display vector pMECS with diversity of 2.8 x 10(9). Based on phage display technology and periplasmic extraction ELISA, H7N2-specific VHH antibodies were successfully isolated. According to a pairing test, two VHH antibodies (Nb79 and Nb95) with good thermal stability and specificity can recognize different epitopes of H7N2 virus. The capture antibody (Nb79) was biotinylated in vivo, and the detection antibody (Nb95) was coupled with horseradish peroxidase (HRP). Based on biotin-streptavidin interaction, a novel sandwich immune ELISA was performed to detect H7N2. The immunoassay exhibited a linear range from 5 to 100 ng/ml. Given the above, the newly developed VHH antibody-based double sandwich ELISA (DAS-ELISA) offers an attractive alternative to other diagnostic approaches for the specific detection of H7N2 virus. (C) 2015 Elsevier Inc. All rights reserved.
资助项目	Jiangsu Nanobody Engineering and Research Center of China[2014-10] ; Program for New Century Excellent Talents in University[NCET-20130127] ; National Natural Science Foundation of China[31271365] ; National Natural Science Foundation of China[31471216] ; Programs for Chang Jiang Scholars and Innovative Research Team in University[IRT1119]
WOS关键词	SENSITIVE DETECTION ; A VIRUS ; NANOBODY ; PCR ; IMMUNOASSAY ; H5N1 ; CONSTRUCTION ; INFECTION ; TOXIN ; ASSAY
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
语种	英语
出版者	ACADEMIC PRESS INC ELSEVIER SCIENCE
WOS记录号	WOS:000373968700014
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/276063]
专题	药理学第一研究室
通讯作者	Wan, Yakun
作者单位	1.Guangxi Med Univ, Guangxi Key Lab Biol Targeting Diag & Therapy Res, Nanning 530021, Guangxi, Peoples R China; 2.Chinese Acad Sci, Shanghai Inst Mat Med, CAS Key Lab Receptor Res, Shanghai 201203, Peoples R China; 3.Guangxi Med Univ, Natl Ctr Int Res Targeting Diag & Therapy, Nanning 530021, Guangxi, Peoples R China; 4.Guangxi Med Univ, Dept Immunol, Nanning 530021, Peoples R China 5.Southeast Univ, Inst Life Sci, Nanjing 210096, Jiangsu, Peoples R China;
推荐引用方式 GB/T 7714	Gong, Xue,Zhu, Min,Li, Guanghui,et al. Specific determination of influenza H7N2 virus based on biotinylated single-domain antibody from a phage-displayed library[J]. ANALYTICAL BIOCHEMISTRY,2016,500:66-72.
APA	Gong, Xue,Zhu, Min,Li, Guanghui,Lu, Xiaoling,&Wan, Yakun.(2016).Specific determination of influenza H7N2 virus based on biotinylated single-domain antibody from a phage-displayed library.ANALYTICAL BIOCHEMISTRY,500,66-72.
MLA	Gong, Xue,et al."Specific determination of influenza H7N2 virus based on biotinylated single-domain antibody from a phage-displayed library".ANALYTICAL BIOCHEMISTRY 500(2016):66-72.