Anti-Infliximab (Remicade®)Free Drug/ADA Dual ELISA Kit

Enzyme-linked immunosorbent assay for the semi-quantitative determination of total and free antibodies to infliximab in serum and plasma. Free and Total Anti-Drug Antibody (ADA) monitoring gained high importance along with measuring free drug from the patient samples. In order to make these measurements simple yet more informative, we have produced our ELISA Kit to measure Free and Total anti-drug antibodies to Infliximab simultaneously on the same plate.

The Assay Genie Antibody to infliximab (Remicade®) ELISA 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 infiliximab specific antibody-infiliximab immune complex is provided by adding dissociation buffer.

After transferring dissociation mix to the plate, antibodies to infliximab (ATI) separated from infiliximab in patient serum/ plasma samples are captured by the drug infliximab (Remicade®) coated on the wall of the microtiter wells and peroxidase-labelled specific conjugate. 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 colour is directly proportional to the concentration of ATI in sample.

Assay Genie Duo ELISA kit can be also used as a semi-quantitative (screening) test in free ADA determination without dissociation and neutralization steps. Peroxidase-labelled specific conjugate and diluted serum / plasma samples are transferred simultaneously to the infliximab-coated plate and antibodies to infliximab (ATI) in patient serum/ plasma samples are captured by the drug infliximab (Remicade®) coated on the wall of the microtiter wells and peroxidase-labelled specific conjugate. 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 colour is directly proportional to the concentration of ATI in sample.

Infliximab (Remicade®) is a chimeric monoclonal antibody and used to treat autoimmune disorders. Infliximab reduces the amount of active human tumour necrosis factor alpha (hTNF alpha) in the body by binding to it and preventing it from signalling the receptors for TNF alpha on the surface of various cell types. TNF alpha is one of the key cytokines that triggers and sustains inflammatory reactions

Infliximab (Remicade®) is used for the treatment of psoriasis, Crohn’s disease, ankylosing spondylitis, psoriatic arthritis, ulcerative colitis, and approved by FDA.

One of the major concern, despite of its wide usage, is potential development of anti-infliximab antibodies (ATI) which in turn may interfere with infliximab (Remicade®) efficacy as mainly judged by observing the relapse of signs and symptoms of disease and necessitate dose-escalation or potentially ending up the treatment.

In this context, demonstration of anti-infliximab antibodies during treatment with infliximab (Remicade®) has a major concern and monitoring for the presence and/or quantitation of specific antibodies during clinical trials is an important issue for follow up of the treatment regimens. With the Assay Genie Duo ELISA Kit infiliximab-specific antibodies (bound or free) can be detected simultaneously.