Anti-Envelope protein [5A]
Catalogue Number: AB03416-23.0-BT-ABA
| Manufacturer: | Vector Laboratories, Inc (ABA) |
| Type: | Recombinant Monoclonal |
| Alias: | E protein; Envelope protein E; Genome polyprotein |
| Shipping Condition: | Blue Ice |
| Unit(s): | 1 mg |
| Host name: | Rabbit |
| Clone: | 5A |
| Isotype: | IgG |
| Immunogen: | 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. |
| Application: | ELISA, IP, FA, NT, InVivoA, Crstapy |
Additional Text
Purification
Purified
Uniprot ID
P03314
Antibody Clonality
Recombinant Monoclonal
Gene ID
1502173
Gene Name
POLY
Storage Note
Store at 4⁰C for up to 3 months. Note, this antibody is provided without added preservatives, it is therefore recommed this antibody be handled under sterile conditions. For longer storage, aliquot and store at -20⁰C.
Application Notes
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).
Short Description
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.
AB03416-23.0-BT Datasheet
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