Abstract: Inflammatory factors and cytokines are an important indicator of the immune response towards new cell therapies and other biologics. Based on its original inflammatory triggers, the diversity of inflammatory factors can vary widely. As such, clinical study samples are focused on quantifying the level of cell factors (such as cytokines) after administration of novel therapeutics. However, samples obtained from clinical trials are subject to several limitations such as sample type, amount, and volume. As such, only a select number of factors can be quantified at a single time. Through traditional methods, this greatly limits the number of studies that can be performed on a single sample due to incompatibilities with the sample matrix, sample volume consumed per test, and total number of samples to be tested. Another challenge is also the resources available at clinical study centers across the world. Whereas top-of-the-line flow cytometers may not be readily available, utilizing cost-effective instruments can reduce the entry barrier and information gathered. To overcome these challenges, a simple, multiplexed flow cytometry-based assay platform was developed to meet the stringent conditions to operate in clinical studies. This platform requires only two fluorescent activation channels for operation: the APC channel dedicated towards separating each marker by its fluorescent intensity and the PE channel used to quantify the concentration. Unlike traditional multiplexed assays that require multiple fluorescent channels, this platform unlocks the upper boundary of quantifying over 10 factors simultaneously within a single 15 µL sample injection and a limit of detection of 1 pg/mL. By minimizing the instrument requirements, sample volume, and enabling a flexible, 10+ multiplexing panel capability, clinical study samples across the globe can obtain a wider variety of information from each patient sample and improve clinical research.
