CloneID: 5A
Antigen Long Description: The original antibody was generated by phage display method. Two antibody-phage libraries were constructed by repertoire cloning from yellow fever patients, subsequently pooled and panned with glycerol-purified non-inactivated YFV-17D virions.
Buffer Composition: PBS only.
Chimeric Use Statement: This full-length chimeric rabbit antibody was made using the variable domain sequences of the original Human ScFv format for improved compatibility with existing reagents assays and techniques.
Uniprot Accession No.: P03314
Specificity Statement: The antibody recognises a quaternary epitope on the envelope protein. The antibody does not bind to E proteins of ZIKV, DENV, or WNV.
Application Notes (Clone): The specificity of the scFv format of the antibody was confirmed by ELISA analysis. The antibody showed 50% and 100% neutralizing activity in plaque reduction neutralization assay (PRNT) using Vero cells against YFV strains 17D-204-WHO and Asibi at concentrations of approximately 1 µg/ml and 10 µg/ml, respectively. The antibody neutralized wild-type YFV strains of genotypes West Africa I (Nigeria 1987) and II (Asibi) and East/Central Africa (CAR 1986, Ethiopia 1961) with comparable efficiency in a PRNT on PS cells. Competitive ELISA showed that the scFv fragment competed for binding to YFV-17D virions with Ab02826. An immunoprecipitation assay with radiolabeled, purified YFV-17D virions was performed. The scFv fragment was found to precipitate a 55-kDa protein, corresponding to the molecular weight of the envelope glycoprotein (Daffis et al., 2005; PMID: 15919103). The structure of the scFv fragment of the antibody in complex with the E protein was determined. Surface plasmon resonance (SPR) experiments showed that the antibody bound to sE proteins from both YFV-17D and YFV-China with high affinities (KD values are 13.5 nM and 9.7 nM, respectively). The in vitro neutralizing activity for YFV was assessed using a modified fluorescence-activated cell sorting (FACS)-based assay in Vero cells. The antibody displayed extremely high neutralization activity against both YFV strains (IC50, 15 ng/mL and 6 ng/mL for YFV-China and YFV-17D, respectively). Mice treated with the antibody were completely protected against YFV-17D infection. Pre- and post-attachment neutralization assays were carried out in BHK-21 cells. In both cases the antibody efficiently neutralized the virus ((IC50= 19 ng/mL and IC50= 8 ng/mL respectively). Finally, the antibody partially blocked pH-dependent fusion of YFV-17D with liposomes (IC50, 19 ng/mL) (Lu et al., 2019).
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