United Kingdom (UK)
Recombinant monoclonal antibody to Nerve growth factor. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma mab 911.
UniProt Accession Number of Target Protein: P01138
Alternative Name(s) of Target: Beta-nerve growth factor; Beta-NGF
Specificity: The antibody is specific for NGF. The NGF epitope is composed of β-strand segments from both NGF monomers. NGF is involved primarily in the growth, as well as the maintenance, proliferation, and survival of nerve cells (neurons).
Application Notes: The specificity of the original format of the antibody to NGF was confirmed by ELISA analysis (EC50 = 0.37 nM). The antibody was evaluated for its ability to block the binding of hNGF to the TrkA and p75 NGF receptors in various in vitro assays, such as blocking of TrkA autophosphorylation and blocking of NGF-dependent survival of dorsal root ganglion sensory neurons. The antibody was a potent blocker of all activities (Hongo et al., 2000; PMID: 10952410). In vivo administration of the antibody significantly reduces bone cancer pain behaviors (Sevcik et al., 2005; PMID: 15836976 and Halvorson et al., 2005; PMID: 16230406 and Buehlmann et al., 2019; PMID: 30161041) and fracture pain-related behaviors (Koewler et al., 2007; PMID: 17638576). The structure of Nerve Growth factor in complex with the Fab fragment was determined. Library Scanning Mutagenesis method was used to convert the original antibody in the humanized antibody tanezumab. The antibody bound human and murine NGF with high affinity (KD ~10 nM). Tanezumab and the original antibody blocked both TrkA and p75NTR binding to NGF and inhibited NGF-dependent neuron survival. Tanezumab inhibited NGFdependent survival with an IC50 of 15 pM; under the same conditions, 911 inhibited NGF with an IC50 of 400 pM (La Porte et al., 2014; PMID: 24830649). Preventative and therapeutic treatment with the antibody significantly prevented, or reversed, MIA-induced pain behaviour in osteoarthritis (Xu et al., 2016; PMID: 27208420).
Antibody first published in:
Hongo et al. Antibody binding regions on human nerve growth factor identified by homolog- and alanine-scanning mutagenesis. Hybridoma. 2000 Jun;19(3):215-27. PMID:10952410
Note on publication:
The original paper describes the generation and characterization of a panel of monoclonal antibodies.
Western blot using anti-Nerve growth factor antibody mab 911 (Ab03301). Human brain cerebellum tissue lysate (35µg protein in RIPA buffer) was resolved on an SDS-PAGE gel, and blots were probed with the monoclonal antibody mab 911 (Ab03301-23.0) at 1µg/ml before detection using an anti-mouse secondary antibody. A primary incubation of 1 hour was used, and protein was detected by chemiluminescence.
Immunofluorescence staining of NIH3T3 cells with anti-Nerve growth factor antibody mab 911 (Ab03301). Immunofluorescence analysis of paraformaldehyde fixed NIH3T3 cells, permeabilized with 0.15% Triton, on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of mab 911 (Ab03301-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Panels show, from left-right, top-bottom, Ab03301-23.0, DAPI, merged channels, and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.
Flow cytometry using anti-Nerve growth factor antibody mab 911 (Ab03301). Paraformaldehyde fixed NIH3T3 cells permeabilized with 0.5% Triton were stained with the anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of mab 911 (Ab03301-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000, and the cells were analyzed using a FACSCanto flow-cytometer.
Western blot using anti-Nerve growth factor antibody mab 911 (Ab03301). Human brain cerebellum tissue lysate (35µg protein in RIPA buffer) was resolved on an SDS-PAGE gel, and blots were probed with the monoclonal antibody mab 911 (Ab03301-23.0) at 1µg/ml before detection using an anti-mouse secondary antibody. A primary incubation of 1 hour was used, and protein was detected by chemiluminescence.