Construction of a novel phage display antibody library against Fasciola hepatica, and generation of a single-chain variable fragment specific for F. hepatica cathepsin L1.
Amino Acid Sequence
Animals
Antibodies, Helminth
/ immunology
Antibodies, Monoclonal
/ immunology
Antigens, Helminth
/ immunology
Cathepsins
/ immunology
Cell Surface Display Techniques
Cloning, Molecular
Electrophoresis, Agar Gel
Fasciola hepatica
/ immunology
Gene Library
Immunoblotting
Immunoglobulin Variable Region
/ chemistry
Peptide Library
Polymerase Chain Reaction
Rats
Rats, Inbred Lew
Single-Chain Antibodies
/ genetics
Cathepsin
Fasciola hepatica
Phage display
Single-chain variable fragment (scFv)
Journal
Experimental parasitology
ISSN: 1090-2449
Titre abrégé: Exp Parasitol
Pays: United States
ID NLM: 0370713
Informations de publication
Date de publication:
Mar 2019
Mar 2019
Historique:
received:
04
07
2018
revised:
02
12
2018
accepted:
10
02
2019
pubmed:
15
2
2019
medline:
28
3
2019
entrez:
15
2
2019
Statut:
ppublish
Résumé
Phage display technology to produce recombinant monoclonal antibodies or antibody fragments permits the identification of sought after antibodies in short time frames at low cost along with direct and rapid selection for antibody characteristics. Monoclonal antibodies can facilitate the identification and characterisation of parasite molecules that function at the host-parasite interface to help understand at the molecular level the biology of the parasite and disease progression, which often leads to new drug targets, diagnostic antigens or vaccine candidates. The trematode Fasciola hepatica is an important veterinary and human parasite. In this work, we infected rats with F. hepatica and amplified the generated antibody repertoire to produce a single-chain variable fragment (scFv) phage display library. The library was used to identify a scFv that recognises cathepsin L1, a major component of the adult parasites excretory/secretory material and an important vaccine candidate. This is the first report of the construction of a phage display antibody library from a F. hepatica infected host, and also the first instance such a library has been used to identify an affinity-matured monoclonal antibody fragment that binds to a F. hepatica antigen. The scFv library and methods detailed should facilitate future research characterising F. hepatica antigens.
Identifiants
pubmed: 30763571
pii: S0014-4894(18)30325-4
doi: 10.1016/j.exppara.2019.02.001
pii:
doi:
Substances chimiques
Antibodies, Helminth
0
Antibodies, Monoclonal
0
Antigens, Helminth
0
Immunoglobulin Variable Region
0
Peptide Library
0
Single-Chain Antibodies
0
Cathepsins
EC 3.4.-
cathepsin L1, Fasciola hepatica
EC 3.4.22.-
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
87-94Informations de copyright
Copyright © 2019 Elsevier Inc. All rights reserved.