United Kingdom (UK)
Recombinant monoclonal antibody to DR4. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma mAY4.
UniProt Accession Number of Target Protein: O00220
Alternative Name(s) of Target: CD261; Death receptor 4; Tumor necrosis factor receptor superfamily member 10A; TNF-related apoptosis-inducing ligand receptor 1; TRAIL receptor 1; TRAILR1; TRAIL-R1; TRAILR 1; TNFRSF10A; APO2
Immunogen: mAb mAY4 was generated by immunising BALB/c mice with the ECD (residues 1-215) of human DR4. Splenocytes were obtained from immunised mice and fused with murine Sp2/O myeloma cells. mAY4 was cloned by RT-PCR and reformatted as an scFv.
Specificity: mAY4 was shown to bind DR4 by ELISA (Sung et al, 2009). These experiments also showed that mAY4 did not react with other homologous death receptors, including DR5, decoy receptor (DcR) 1, DcR2, FAS receptor, and mTR (a mouse ortholog of human DR4 and DR5). SPR showed an affinity of 7.1 nM for interaction with DR4. mAY4 bound to a different epitope on DR4 than the physiological ligand TRAIL. DR4 is a pro-apoptotic receptor expressed on all cells. It represents a promising target for anti-cancer therapeutic agents.
Application Notes: mAY4 was shown to bind DR4, but not other related homologs, using ELISAs and SPR (Sung et al, 2009). The mAb and scFv-reformatted version were also used in cell viability assays for assessment of tumouricidal activity. More recently, the mAY4 CDRs were humanised and expressed as an scFv-reformatted antibody fragment (Lee et al, 2010), although the humanised version had a 10-fold lower affinity for DR4.
Antibody first published in:
Sung et al. A novel agonistic antibody to human death receptor 4 induces apoptotic cell death in various tumor cells without cytotoxicity in hepatocytes. Mol Cancer Ther. 2009 Aug;8(8):2276-85. PMID:19638452
Note on publication:
Describes the generation of mAY4, a murine antibody against human DR4. The mAb, and a scFv-reformatted version, were shown to promote DR4-mediated apoptotic activity and prevent tumour growth of human NSCLC tumours established in mice.
Flow cytometry using the Anti-DR4 antibody mAY4 (Ab00953). Paraformaldehyde fixed human peripheral blood monocytes permeabilized with 0.5% Triton were stained with anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of mAY4 (Ab00953-23.0, 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 cells analyzed using a FACSCanto flow-cytometer.