Three new assays for rituximab based on its immunological activity or antigenic properties: analyses of sera and plasmas of RTX-treated patients with chronic lymphocytic leukemia and other B cell lymphomas
Abstract
Rituximab (RTX) is a monoclonal antibody which targets CD20 and is approved for treatment of non-Hodgkin's lymphoma (NHL), with an approximate 50% overall response rate among NHL patients. Accurate determination of RTX concentrations in patient plasmas is important for proper dosing of patients and for correlating RTX concentrations with clinical responses. There is currently no assay available for RTX which utilizes easily obtainable commercial reagents. Therefore, we sought to develop such an assay, and in this report we describe three new assays for RTX concentration. One assay, based on flow cytometry, quantitates immunologically active RTX based on its ability to bind to CD20 on Raji cells. Two other methods, based on flow cytometry and ELISA, measure RTX based on its antigenic properties. The assays are accurate, in good agreement with one another, and can all measure RTX concentrations as low as approximately 1 microg/ml in both sera and plasmas. Use of these assays reveals that chronic lymphocytic leukemia (CLL) patients receiving RTX treatment have lower plasma RTX concentrations than patients with other B cell lymphomas at all times over the usual 4-week course of therapy. The level in CLL plasmas often declines to <1...Continue Reading
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