United Kingdom (UK)
Recombinant monoclonal antibody to FedF. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from phage display NbFedF9.
UniProt Accession Number of Target Protein: Q47212
Alternative Name(s) of Target: F18 fimbrial adhesin AC; F18 fimbrial adhesin
Immunogen: A Llama was immunised 6 times with purified FedF residues 15-165 over a period of 6 weeks. Lymphocytes were extracted and used to prepare cDNA which was used as a template fthe VHH. These were then transfected into E. coli and the antibodies selected by phage display. This was then subjected to ELISA to confirm the specificity of the antibodies.
Specificity: Specific to the N terminal lectin domain as this is the domain represented in the 15-165 fragment. It has an affinity for FedF(15–165) of around 1.5 nM (Moones et al, 2014).
Application Notes: It has been shown that this antibody can inhibit the binding of FedF to piglent enterocytes. The structure of this antibody binding to FedF has been resolved to a resolution of 1.5A. This shows that the mechanism of inhibition of FedF binding to epithelial cell surfaces is by competing at the carbohydrate binding site. Both MST and SPR have been used to analyse binding affinities (Moones et al, 2014).
Antibody first published in:
Moonens et al Nanobody Mediated Inhibition of Attachment of F18 Fimbriae Expressing Escherichia coli PLoS One. 2014; 9(12): e114691. PMID:25502211
Note on publication:
Discusses the generation of this antibody, its crystallisation and the method of netralisation of FedF