Generation of a single-chain variable fragment phage display antibody library from naive mice panned against Fasciola hepatica antigens

被引:5
作者
Norbury, Luke J. [1 ]
Basalaj, Katarzyna [1 ]
Baska, Piotr [2 ]
Zawistowska-Deniziak, Anna [1 ]
Kalinowska, Alicja [1 ]
Wilkowski, Przemyslaw [1 ]
Wesolowska, Agnieszka [1 ]
Wedrychowicz, Halina [1 ]
机构
[1] Polish Acad Sci, Witold Stefanski Inst Parasitol, Twarda 51-55, PL-00818 Warsaw, Poland
[2] Warsaw Univ Life Sci SGGW, Fac Vet Med, Div Pharmacol & Toxicol, Ciszewskiego 8, PL-02786 Warsaw, Poland
关键词
Fasciola hepatica; Phage display; Cathepsin; Single-chain variable fragment (scFv); IMMUNE-RESPONSE; EXCRETORY/SECRETORY PRODUCTS; DIFFERENTIAL EXPRESSION; MONOCLONAL-ANTIBODIES; CATHEPSIN L3; PROTEIN; REPERTOIRE; CONSTRUCTION; ENVIRONMENT; DIAGNOSTICS;
D O I
10.1016/j.exppara.2019.107737
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
Monoclonal antibodies have a wide range of applications in basic and applied research as well as in the medical and pharmaceutical industries. Phage display antibody libraries offer an alternative to hybridoma technology for the generation of monoclonal antibodies and can be applied to high-throughput screening and facilitate the generation of novel antibodies. Despite their utility in several fields of research there has been limited application of antibody libraries in the study of trematode parasites. Fasciola hepatica causes considerable loss to the agriculture sector and is also a human pathogen. The parasite's excretory/secretory material contains numerous molecules that facilitate its invasion and survival within the mammalian host, including cathepsin B and L proteases. F. hepatica cathepsin B2 is expressed during the initial weeks of infection and has suspected roles in immune evasion and as a digestive enzyme in the parasite's gut; it is considered a good target for vaccination or therapeutic inhibitors. In this study, we produced a single-chain variable fragment (scFv) phage display library from naive mice. The library was used to identify several scFv that can bind to antigens from adult F. hepatica homogenate, and a scFv that can bind to F. hepatica cathepsin B2. The results highlight the potential applicability of such a library to facilitate the study of F. hepatica and other parasites. This is the first report of the application of a naive phage display antibody library to the study of F. hepatica.
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页数:7
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