CloneID: 3F1H10
Antigen Long Description: 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 3F1H10 was selected from a subsequent fusion after three more months of continued immunization.
Origin Pub PMID: 10938729
Buffer Composition: PBS with 0.02% Proclin 300.
Available Custom Conjugation Options: AP, HRP, Fluorescein, APC, PE, Biotin Type A, Biotin Type B, Streptavidin, FluoroProbes 647H, Atto488, APC/Cy7, PE/Cy7
Uniprot Accession No.: P27662
Specificity Statement: 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.
Application Notes (Clone): The neutralizing activity of the original format of this antibody (mouse IgG1) was measured at the cell culture level in a plaque neutralization test (50% PNT). This antibody was administered to rainbow trout via intraperitoneal injection to evaluate its passive immunization protective effect and efficacy against Egtved virus infection. This antibody was used as the primary antibody in immunoblotting (IB) to detect the viral glycoprotein. This antibody, conjugated with fluorochromes like FITC or TRITC, was employed to visualize and detect Egtved virus-infected cells. This antibody, which was highly neutralizing in vitro, also provided the highest degree of protection in vivo of the antibodies tested (Lorenzen et al., 1990; PMID: 1690259). This antibody's neutralizing activity against different VHSV isolates was determined by 50% PNT; This antibody's original format neutralized type I isolates VHSV (I-F1 and I-92) well, but higher concentrations were needed to neutralize type II isolates and even higher concentrations were required to neutralize type III isolates. An 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 3.8 nM for the immobilized G protein of VHSV strain I-F1 and a KD of 64 nM for the immobilized G protein of VHSV strain I-92 (Lorenzen et al., 2000; PMID: 10938729). Monovalent versions of this antibody, in the form of Fab and recombinant single-chain antibody fragments (scAbs), were able to neutralize VHSV, indicating that the Fc moiety and divalency of the antibody molecules are not necessary 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 antibody, while the kas were similar. The increased kd was accompanied by decreased neutralizing activity. The KD measured against immobilized G protein of VHSV strain DK-F1 was 3.8 nM for the parent antibody and 124 and 241 nM for the daughter Fab and scAb, respectively (Cupit et al., 2001; PMID: 11682124).
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