Enhanced cytokine detection by a novel cell culture-based ELISA

Production of some cytokines, such as IL-4 and IL-10, often occurs at low levels and is difficult to detect by standard ELISA techniques. In many cases the level of detection is at or near to the limits of sensitivity of the assay due either to minimal synthesis and/or cytokine consumption. In an effort to enhance the quantitation of weakly detected cytokines we have developed a unique cell culture-capture ELISA. Lymphocytes are incubated in an anti-cytokine antibody coated ELISA plate for the last 6 hours of a 24 hour in vitro activation period. Use of this cell culture capture method consistently enhanced detection of several T cell cytokines compared to conventional ELISA techniques. Moreover, this technique was found to enhance detection without altering the rate of cytokine secretion which occurred prior to the cell culture capture period. Thus, the cell culture capture ELISA may be useful for detection of a variety of cytokines which are produced at low levels and have traditionally been difficult to quantify.