Eculizumab ELISA (SOLIRIS)

This Eculizumab ELISA is for determination of Eculizumab in human serum, plasma and cell culture supernatant.

Assay Background: Eculizumab is used to treat paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), and neuromyelitis optica. Eculizumab, sold under the trade name Soliris, binds to terminal complement component 5 (C5). C5 is part of activation of host cells, helping attract pro inflammatory immune cells. Inhibition of the complement cascade by inhibition of C5 cleavage, impeding inflammation and cell destruction.

Principle of the assay: The method employs the quantitative sandwich enzyme immunoassay technique. Antibodies to Eculizumab are pre-coated onto microwells. Samples and standards are pipetted into microwells and human Eculizumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated anti- Eculizumab antibody is pipetted and incubated. After washing microwells in order to remove any non-specific binding, the ready to use substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Eculizumab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.