Profiling 115 proteins in colon and duodenal organoid supernatants by multiplex immunoassay 3082

Abstract Description MILLIPLEX® PLEXpedition is a configurable multiplex immunoassay kit that can simultaneously measure up to 115 targets including cytokines, chemokines, growth factors, matrix metalloproteinases, and biomarkers of bone health, metabolism, and cardiovascular disease. These 115 proteins were initially profiled in a diverse collection of human biofluids, including serum, plasma, cerebrospinal fluid, urine, milk, synovial fluid, and bronchoalveolar lavage (BAL). To evaluate the utility of this kit in ex vivo research models, we assessed 3dGRO® Gastrointestinal Organoids cultured in supplemented L-WRN Conditioned Medium. Supernatants were collected after 48 and 72 hours from matched colon and duodenal organoid cultures from a 21-year-old donor. The vast majority of the 115 analytes were detectable in at least one of the supernatant samples and were increased in the organoid supernatants relative to the conditioned media control. However, proteins exogenously added to the organoid culture media (Insulin, EGF) were above the upper limit of quantification. Most analytes were increased at the 72-hour timepoint relative to the 48-hour timepoint. A subset of analytes exhibited differential expression based on the tissue of origin, displaying relative increases in either the colon or duodenal organoid supernatants. In summary, this study demonstrates how a multiplex immunoassay can be applied to organoid research and used to measure >100 proteins in organoid supernatants. Topic Categories Technological Innovations in Immunology (TECH)