జర్నల్ ఆఫ్ ఎనలిటికల్ & బయోఅనలిటికల్ టెక్నిక్స్

నైరూప్య

Development of Improved Sandwich Elisa for the In Vitro Detection of Inhibitors of the TNF-TNFR₁ Interaction

Jessica M Davis

The over-expression of tumor necrosis factor alpha (TNF-alpha) has been associated with various diseases, particularly autoimmune diseases such as rheumatoid arthritis and Crohn’s disease. Some biologic therapies for these diseases specifically target this cytokine, sequestering it, and preventing its interaction with its 55 kD receptor (TNFR 1 ). While these therapies have proven the validity of this approach, they are large proteins that require intravenous administration. Small molecule inhibitors of this interaction are highly desirable and some have been developed. However, such inhibitors have not been reported to have successfully completed clinical trials. One prohibition to this approach is the current state of in vitro testing of such molecules. Assays have been developed that use radio labeled or biotin labeled TNF-alpha. In vivo methods exist, but are cost prohibitive to screening a large selection of molecules. Reported here-in is an inexpensive, highly robust, sensitive assay for inhibitors of the TNF- alpha interaction with TNFR 1 that does not require modification of TNF-alpha. The simplicity of this assay should allow for increased investigation into inhibiting this important interaction.