Construction, screening and identification of a phage display antibody library against the Eimeria acervulina merozoite

被引:10
|
作者
Zhao, Yuelan [2 ]
Amer, Said [3 ]
Wang, Jianyong [2 ]
Wang, Chengmin [4 ]
Gao, Ying [1 ]
Kang, Guiying [5 ]
Bao, Yongzhan [1 ]
He, Hongxuan [4 ]
Qin, Jianhua [1 ]
机构
[1] Agr Univ Hebei, Coll Anim Sci & Technol, Baoding 071001, Peoples R China
[2] Agr Univ Hebei, Coll Chinese Tradit Vet Sci, Dingzhou 073000, Peoples R China
[3] Fac Sci, Dept Zool, Kafr Al Sheikh 33516, Egypt
[4] Chinese Acad Sci, Inst Zool, Key Lab Anim Ecol & Conservat Biol, Natl Res Ctr Wildlife Borne Dis, Beijing 100101, Peoples R China
[5] Inner Mongolia Univ Nationalities, Coll Anim Sci & Technol, Tongliao 028000, Peoples R China
基金
中国国家自然科学基金;
关键词
Eimeria acervulina; Merozoite; Phage display; Single-chain antibody; Chicken; VACCINE DEVELOPMENT; COCCIDIOSIS; CHICKEN; SPOROZOITE; FRAGMENTS; POULTRY; SELECTION; PARASITES; IMMUNITY; ANTIGEN;
D O I
10.1016/j.bbrc.2010.02.063
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A single-chain antibody library against Eimeria acervulina merozoites was constructed by phage display approach. Antibody-displaying phage was selected in four panning rounds against cryopreserved E. acervulina merozoites. Five clones were randomly selected from the fourth panning round, and their nucleotide sequences were aligned and compared to mouse germ-line sequences. Soluble antibody was produced in a non-suppressor Escherichia coli strain, purified by protein A affinity chromatography, and characterized by Western-blotting. Immunofluorescence assay showed localization of the produced recombinant antibody fragment on the surface E. acervulina merozoites. These resultant antibody fragments showed high specificity and binding capacity for soluble antigens and intact fixed merozoites which seems promising as diagnostic, therapeutic and/or vaccine tools against coccidiosis. Crown Copyright (C) 2010 Published by Elsevier Inc. All rights reserved.
引用
收藏
页码:703 / 707
页数:5
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