Cat. No.
MABL-943
Application
IP, SPR, Block, ELISA, FC, IHC
Isotype
Engineer antibody
Species Reactivity
Cynomolgus, marmoset monkey, Human
Clone No.
5D12
From
Recombinant Antibody
Specificity
ch5D12 binds specifically to recombinant and native human CD40 ectodomain and has been shown to also bind CD40 in cynomolgus and marmoset monkeys. The antibody requires residues within D1 and D1/B2 for binding (Hager et al, 2003) (D1 domain is crucial for CD40L binding). Binding characteristics are very similar between humanized ch5D12 and mu5D12. CD40 is a glycoprotein of the TNFR superfamily and is expressed on all mature B cells, dendritic cells, activated monocytes, some endothelial cells and some epithelium including the thymus. CD40 binds to CD40L, and this interaction is involved in B-cell activation and proliferation, antigen-presenting cell (APC) activation, initiation of antigen-specific T-cell responses, immunoglobulin production, activation of effector macrophages isotype switching, homotypic adhesion and rescue from apoptosis.
Alternative Names
TNFRS5;Tumor necrosis factor receptor superfamily member 5; B-cell surface antigen CD40; Bp50; CD40L receptor; CDw40
UniProt
P25942
Immunogen
Anti-CD40 mAb 5D12, was generated by immunizing mice with sf9 insect cells expressing recombinant human CD40 and selected for the ability to bind EBV-immortalized human B cells. The variable regions from mu5D12 were cloned and used to construct chimeric humanized IgG4 5D12.
Application Notes
ch5D12 is an antagonist of the CD40-CD40L pathway. ch5D12 binds to B cells and monocytes, but not to peripheral blood CD4+ and CD8+ cells. ch5D12 was tested in vivo using a marmoset EAE (experimental autoimmune encephalomyelitis) model (recombinant myelin oligodendrocyte glycoprotein (MOG) was used to induce EAE) - model for human multiple sclerosis. ch5D12 was able to prevent development of the symptoms of the disease during the treatment period, reduce the anti-MOG antibody response and reduce the formation of inflammatory lesions in the CNS. ch5D12 binding and specificity can be detected and measured by FC, SPR, ELISA and IP, and CD40+ cells can be stained by the antibody in IHC.
Antibody First Published
Boon et al. Preclinical assessment of anti-CD40 Mab 5D12 in cynomolgus monkeys Toxicology. 2002 May 15;174(1):53-65. PMID:11972992
Note on publication
Describes the creation of a chimeric human IgG4 version of mu5D12 to enhance the in vivo half-life and reduce the immunogenicity of the antibody capable of antagonising the CD40-CD40L interaction.
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.
