Alemtuzumab ELISA (LEMTRADA)

Intended Use:This Alemtuzumab ELISA is a rapid and easy method for the quantitative determination of Alemtuzumab in human serum, plasma and cell culture supernatan Assay Background:Alemtuzumab (marketed as Campath, MabCampath or Campath-1H and currently under further development as Lemtrada) is a monoclonal antibody used in the treatment of chronic lymphocytic leukemia (CLL), cutaneous T-cell lymphoma (CTCL) and T-cell lymphoma. It is also used in some conditioning regimens for bone marrow transplantation, kidney transplantation and Islet cell transplantation. Alemtuzumab binds to CD52, a protein present on the surface of mature lymphocytes, but not on the stem cells from which these lymphocytes are derived. After treatment with Alemtuzumab, these CD52-bearing lymphocytes are targeted for destruction. Alemtuzumab is used as second-line therapy for CLL. It was approved by the US Food and Drug Administration for CLL patients who have been treated with alkylating agents and who have failed fludarabine therapy. It has been approved by Health Canada for the same indication, and additionally for CLL patients who have not had any previous therapies. Assay Principle:The method employs the quantitative sandwich enzyme immunoassay technique. Antibodies to Alemtuzumab are pre-coated onto microwells. Samples and standards are pipetted into microwells and human Alemtuzumab present in the sample are bound by the capture antibody. Then, a HRP (horseradish peroxidase) conjugated anti- Alemtuzumab antibody is pipetted and incubated with samples. 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 Alemtuzumab in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.