Simultaneous Quantification of Six Human Cytokines in a Single Sample Using Microparticle-based Flow Cytometric Technology

The study of cytokines has been of great interest to researchers in many scientific disciplines, including cell biology and immunology. Cytokines form a sophisticated network that serves to modulate a myriad of cellular events. Within such a network, and through complex feedback mechanisms, cytokine functions are mostly interdependent. Because of the extreme complexity of the cytokine network, a simultaneous measurement of multiple cytokines in a single sample represents a desirable and effective approach. Several methods are available to measure cytokines and their messenger RNAs. To measure cytokines secreted from cells, researchers commonly use conventional ELISA techniques. ELISA methods are generally cost-effective but restricted to measuring one cytokine at a time. Consequently, the ELISA approach requires not only multiple sample aliquots but also repetitive execution of the procedures for each cytokine of interest. The ability of flow cytometry to simultaneously acquire data for numerous particles and subsequently analyze multiple characteristics for each particle makes it a powerful technique for cell sorting and cell analyses. The application is fundamentally built on the high sensitivity of a flow cytometer to discern characteristics either within or on the surface of a cell. Several investigators have constructed microparticle-based immunoassays that use a flow cytometer …