Additional Text

Gene Name

TNF

Gene ID

7124

Uniprot ID

P01375

Purification

Purified

Antibody Clonality

Recombinant Monoclonal

Storage Note

Store at 4⁰C for up to 3 months. Note, this antibody is provided without added preservatives, it is therefore recommed this antibody be handled under sterile conditions. For longer storage, aliquot and store at -20⁰C.

Short Description

This chimeric mouse antibody was made using the variable domain sequences of the original Human IgG1 format, for improved compatibility with existing reagents, assays and techniques.

Application Notes

The binding affinity of the IgG1 fragment for soluble human TNFa was measured by surface plasmon resonance (Kd= 18 pM). The binfing affinity of the IgG1 fragment for transmembrane human TNFa was measured by radioimmunoassay (Kd= 1890 pM). The original format of the antibody was neutralizing, as shown by its ability to block the binding of human TNFa to recombinant human TNFR1 (IC50= 3.9 ng/mL) and inhibit human TNFa-mediated cell cytotoxicity (IC50= 3.2ng/mL). Differential scanning calorimetry (DSC) was used to evaluate the conformational and thermal stability of the original format of the antibody and the Fab fragment (primary transition temperature of 74.8 and 74.5°C respectively). The cytotoxic effect of TNFa on the human rhabdomyosarcoma cell line KYM-1D4 was used to compare the neutralization of soluble TNFa and tmTNFa by the original format of the antibody (IC50= 6.5 ng/mL and 162 ng/mL respectively). When using primary human umbilical vein endothelial cells (HUVEC) stimulated with soluble TNFa, the IC50 was 3.9 ng/mL. The original format of the antibody has the inherent capacity to induce antibody-dependent and complement-mediated cell lysis. In vivo, the original format of the antibody at doses of 1 and 10 mg/kg significantly delayed disease progression in a mouse model of human TNFa-induced arthritis when compared with untreated mice. The original format of the antibody also significantly reduced histological scores for arthritis severity and cartilage damage, as well as serum levels of pro-inflammatory cytokines and chemokines associated with arthritis (Shealy et al., 2010; PMID: 20519961). The specificity of the original format of the antibody was confirmed by ELISA analysis (US7250165B2). The crystal structure of the Fv fragment of the antibody in complex with TNFa was determined. The crystal structure showed that the antibody exerts its inhibitory effect by preventing binding of TNFR1 and TNFR2 to TNFa by steric hindrance (Ono et al., 2018; PMID: 29575262). The antibody is used as a therapeutic for diseases like rheumatoid arthritis, psoriatic arthritis and ankylosing spondylitis.