Single-chain variable fragment (scFv) antibodies against rotavirus NSP4 enterotoxin generated by phage display

Journal of Virological Methods
Jesús Rodríguez-DíazJavier Buesa

Abstract

The rotavirus non-structural NSP4 protein causes membrane destabilization as well as an increase in intracellular calcium levels in eukaryotic cells and induces diarrhea in young mice, acting as a viral enterotoxin. In this study the phage display technique was used to generate a panel of single-chain variable fragment (scFv) antibodies specific for the NSP4 protein of the human rotavirus strain Wa from a human semi-synthetic scFv library. After several rounds of panning and selection on NSP4 adsorbed to polystyrene tubes, individual scFv were isolated and characterised by fingerprinting and by sequencing the VH and VL genes. The isolated scFv antibodies specifically recognize NSP4 in enzyme immunoassay and in Western blot. Four truncated forms of the NSP4 protein were constructed which allowed us to map the binding region of the selected scFv antibodies to the C-terminal portion of NSP4. The isolated scFv antibodies constitute valuable tools to analyse the mechanisms of NSP4 functions.

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Citations

Mar 25, 2008·Biochemical and Biophysical Research Communications·Esra Acar SoykutIsmail Hakki Boyaci
Nov 28, 2006·Journal of Immunological Methods·Si WuJennifer A Doudna
Aug 27, 2005·Journal of Medical Virology·Jesús Rodríguez-DíazJavier Buesa
Apr 22, 2014·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Yashpal Singh MalikJelle Matthijnssens
May 21, 2016·Proteomics. Clinical Applications·Philipp KuhnMichael Hust
Jul 27, 2021·Frontiers in Cellular and Infection Microbiology·Kristian Daniel Ralph RothMichael Hust

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