Cat. No.
MABL-3215
Application
in vitro, PNT, ELISA
Isotype
Engineer antibody
Species Reactivity
VHSV, Viral hemorrhagic septicemia virus
Clone No.
3F1A2
From
Recombinant Antibody
Specificity
This antibody is specific for the transmembrane envelope glycoprotein (G protein) of viral hemorrhagic septicemia virus (VHSV). The G protein is the only protein known to be present on the surface of the virus particle, and it constitutes less than 10% of the mass of the virus particle which is dominated by N, M1 and M2. This antibody demonstrated similar neutralizing activity to clone 3F1H10, and they were found to interfere with each other's binding reciprocally, which indicates they recognize the same or partially related epitopes.
Alternative Names
G; Spike glycoprotein
UniProt
P27662
Immunogen
The original antibody was generated by immunizing BALB/c mice with Egtved virus by intraperitoneal injections five times over two months, followed by an additional intravenous booster injection on week nine. The hybridoma cell line secreting clone 3F1A2 was selected from a subsequent fusion after three more months of continued immunization.
Application Notes
The neutralizing activity of the original format of this antibody (mouse IgG1) against different VHSV isolates was determined by 50% PNT; it neutralized type I isolates VHSV (I-F1 and I-92) and type II isolate VHSV (II-31) well, but higher concentrations were needed to neutralize type III isolates (III-51 and III-37). ELISA was used to detect and quantify the antibody's binding efficiency to various VHSV strains. The antibody's binding kinetics were evaluated by surface plasmon resonance (SPR) analysis resulting in a KD of 2.4 nM for the immobilized G protein of VHSV strain I-F1 and a KD of 12.1 nM for the immobilized G protein of VHSV strain I-92 (Lorenzen et al., 2000; PMID: 10938729). The Fab and recombinant single-chain antibody fragments (scAbs) versions of this antibody neutralized VHSV, indicating that the Fc moiety and divalency of the antibody molecules are unnecessary for neutralization. However, the neutralizing activity of Fab and scAb fragments was generally lower compared to the parent antibody due to the higher avidity of the divalent parent molecules; the Fab and scAb fragments exhibited a 10-100-fold increase in kd compared to the parent MAb, while the kas were similar. The increased kd was accompanied by decreased neutralizing activity.
Antibody First Published
Lorenzen et al. Three monoclonal antibodies to the VHS virus glycoprotein: comparison of reactivity in relation to differences in immunoglobulin variable domain gene sequences Fish Shellfish Immunol. 2000 Feb;10(2):129-42. doi: 10.1006/fsim.1999.0232 PMID:10938729
Note on publication
The original publication compares three monoclonal antibodies to the glycoprotein of viral hemorrhagic septicemia virus (VHSV) in terms of their reactivity and neutralizing activity, while examining the differences in immunoglobulin variable domain gene sequences and their potential role in antibody functionality.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
