Construction of a Nanobodies Phage Display Library From an Escherichia coli Immunized Dromedary

Document Type: Research Paper

Authors

Livestock and wildlife laboratory, Arid lands Institute, Gabès University, Medenine, Tunisia

Abstract

Background: Diarrhea caused by Escherichia coli is a major cause of morbidity and mortality in young animals. Few treatment options are available, mainly antibiotic therapy increasingly limited by resistance to commonly used drugs.
Objectives: The aim of this work was to develop immunotherapy based on the use of camel VHH antibody fragments, or nanobodies, to target pathogenic E. coli surface antigens.
Material and methods: We immunized a camel with a killed strain we had previously isolated from a diarrheic camel calf and identified as expressing the F17 fimbriae antigen.
Results: The immunized animal developed an anti-E.coli immune response including heavy-chain antibodies. Lymphocytes from this animal were purified and RNA isolated to create a VHH library by phage display with a size of about 109 individual transformants. Panning on live E. coli cells resulted in the isolation of VHH fragments specific to the cell surface antigens.
Conclusion: The identification of these antigens can lead to the development of new diagnostic and therapeutic tools against diarrhea.

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