Anti-Infliximab (REMICADE) Screening ELISA Assay Kit
The Anti-Infliximab (REMICADE) Screening ELISA Assay Kit is For Research Use Only
Size: 12 x 8 wells
Sensitivity: +/-
Incubation Time: 1 hour 20 minutes
Sample Type: Serum, Plasma
Sample Size: 40 µL
Alternative Name: Remicade
Controls Included
Assay Background
Infliximab is a tumor necrosis factor (TNFa) blocker and a chimeric monoclonal IgG1 antibody composed of human constant (75%) and murine variable (25%) regions. Infliximab is produced by a recombinant cell line cultured by continuous perfusion. TN Fa isa key proinflammatory cytokine involved in chronic inflammatory diseases. Its hyperactivity and enhanced signaling pathways can be observed in inflammatory diseases where it activates further pro-inflammatory cascades. By binding to both the soluble subunit and the membrane-bound precursor of TN Fa, infliximab disrupts the interaction of TNFa with its receptors and may also cause lysis of cells that produce TNFa. Infliximab is an IgG1 K monoclonal antibody that binds to soluble and transmembrane forms of TNFa with high affinity to disrupt the pro-inflammatory cascade signaling. Binding of the antibody to TN Fa prevents TN Fa from interacting with its receptors. Infliximab does not neutralize TN Fa (lymphotoxin-a), a related cytokine that utilizes the same receptors as TN Fa. Blocked actions of TN Fa further leads to downregulation of local and systemic pro-inflammatory cytokines (i.e. IL-1, IL-6), reduction of lymphocyte and leukocyte migration to sites of inflammation, induction of apoptosis of TNF-producing cells (i.e. activated monocytes and T lymphocytes), increased levels of nuclear factor-KB, inhibitor, and reduction of reduction of endothelial adhesion molecules and acute phase proteins. Its inhibitory actions on TN Fa was demonstrated in human fibroblasts, endothelial cells, neutrophils, B and T lymphocytes and epithelial cells. Infliximab also attenuates the production of tissue degrading enzymes synthesized by synoviocytes and/or chondrocytes.
Assay Principle
The Anti-Infliximab (REMICADE) Screening ELISA Assay Kit is a sandwich assay for the determination of total and free antibodies against infliximab in serum and plasma samples. During the first incubation period, the separation of infliximab specific antibody- infliximab immune complex is provided by adding dissociation buffer. After transferring dissociation mix to the plate, infliximab antibodies are separated from infliximab in patient serum/plasma samples and they are captured by the drug infliximab coated on the wall of the microtiter wells and horse radish peroxidase (HRP) conjugated probe. After washing away the unbound components from samples, the bound enzymatic activity is detected by addition of tetramethylbenzidine (TMB) chromogen substrate. Finally, the reaction is terminated with an acidic stop solution. The intensity of the reaction color is directly proportional to the concentration of infliximab antibodies in sample.
This kit can be also used as a semi-quantitative test for free anti-drug antibodies determination without dissociation and neutralization steps. Peroxidase labeled specific conjugate and diluted serum/plasma samples are transferred simultaneously to the infliximab-coated plate and antibodies to infliximab in patient serum/ plasma samples are captured by the drug infliximab coated on the wall of the microtiter wells and HRP conjugated probe. After washing away the unbound components from samples, the bound enzymatic activity is detected by addition of TMB chromogen substrate. Finally, the reaction is terminated with an acidic stop solution. The intensity of the reaction color is directly proportional to the concentration of infliximab antibodies in sample.
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