INTRODUCTION

Infliximab (Remicade®) is a chimeric monoclonal antibody against tumour necrosis factor alpha (TNF-α). Infliximab neutralizes the biological activity of TNF-α by binding with high affinity to the soluble (free floating in the blood) and transmembrane (located onthe outer membranes of T cells and similar immune cells) forms of TNF-α, and inhibits or prevents the effective binding of TNF-α with its receptors.

The Infliximab ELISA kit can be used for monitoring infliximab during clinical research and offers the scientists a tool for understanding of the safety and efficacy of the Infliximab and its biosimilars. The assay design utilizes a pair of antibodies that allows the detection of the whole Inflimximab molecule in biological matrices.

PRINCIPLE OF THE ASSAY

This assay employs the sandwich enzyme immunoassay technique. Anti- Infliximab is coated onto a 96 well microplate. Calibrator, quality control samples (if desired) and test samples are pipetted into the appropriate wells. Infliximab present in biological matrices is bound by the immobilized anti- Infliximab antibody. After washing away any unbound substances, enzyme linked anti- Infliximab antibody is added to the wells. This antibody is developed and purified specifically against truncated Remicade® (domain residing in Fc portion of the Infliximab® molecule). The plate is washed to remove any unbound antibody-enzyme reagent and a substrate solution is added to the wells for color development. The color development is proportional to the amount of Infliximab present in test samples. The color development is stopped and the intensity of the color is measured.