Discovery and screening of protein biomarkers with the FirePlex Technology Platform

In patients and animal models, molecular biomarkers are used as indicators of normal and pathogenic processes. In drug discovery and screening pipelines, molecular biomarkers are used to assess the mechanism of action, efficacy, and toxicity of lead compounds. To address the need for rapid and sensitive biomarker quantitation, we have developed the FirePlex® Technology Platform. Utilizing patented hydrogel particles and a three-region encoding design, FirePlex assays allow for true, in-well multiplexing, providing flexible and customizable quantification of miRNA and protein analytes. To facilitate biomarker discovery studies, we offer our standard-throughput, flow cytometry-based assays which enable quantitation of up to 75 miRNA or protein analytes per sample, from only 20 µl of input. These assays demonstrate 5 logs of dynamic range and offer highly sensitive quantitation of analytes in serum, plasma, cell culture supernatant, urine, and CSF without the need for sample processing or RNA isolation. For drug discovery and screening studies, we offer our high-throughput FirePlex (FirePlex®-HT) Immunoassays for quantitation of up to 10 protein analytes per sample from only 6.25 µl biofluid input, in 384-well plate format. FirePlex-HT assays provide 3-4 logs dynamic range, demonstrate 1-100 pg/ml sensitivity, and have been validated in serum, plasma, and cell culture supernatant. The two-step workflow, no-wash assay format, and readout on high-content imagers limit hands-on time and are amenable to automation, thus making FirePlex-HT ideally suited for high-throughput screening studies. In addition, here we introduce our high-throughput miRNA assays for screening miRNA biomarkers in 96-well or 38-4well plate format, with readout on high-content imagers. Here we present data from miRNA and protein profiling studies using the FirePlex platform, and introduce the simplified workflow of the FirePlex-HT® immunoassays with data demonstrating the performance for quantifying key cytokines in multiplex, in biological samples. Together, this novel combination of multiplexed, high-sensitivity assays and bioinformatics tools enables rapid quantitation of protein biomarker signatures in biofluid specimens. Citation Format: Amy Perea, Russell Neuner, Bianca Heinrich, Graeme Doran, Wayne Austin, Conor Rafferty, Matt Camilleri, Michael Tackett, Long To, Elnaz Atabakhsh, James Murray, Daniel Pregibon. Discovery and screening of protein biomarkers with the FirePlex Technology Platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4060.