Category: Assay Development and Screening

1154-E - High-throughput, multiplex quantitation of protein biomarkers using the FirePlex-HT technology Platform

Wednesday, February 7, 2018
11:30 AM - 12:30 PM

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 quantitation of protein biomarkers, we have developed the FirePlex®-HT Immunoassays, which enable multiplex quantitation of up to 10 protein analytes in 384-well plate format.

Utilizing patented FirePlex hydrogel particles and a three-region encoding design, our assays allow for true, in-well multiplexing, providing flexible and customizable quantification of analytes. FirePlex-HT immunoassays use high-performance recombinant matched antibody pairs that reduce cross reactivity between individual analytes, provide up to 3-4 logs dynamic range, and demonstrate 1-100 pg/ml sensitivity. Assays require only 12.5 µl of biofluid sample per well, and have been validated in serum, plasma, and cell culture supernatant. The two-step workflow and no-wash assay format limit hands-on time and are amenable to automation, thus making FirePlex-HT ideally suited for high-throughput screening studies. Assay readout is conducted on high-content imagers and data analysis is performed with the integrated, free-of-charge FirePlex® Analysis Workbench software, thus bypassing the need for dedicated instrumentation or expensive software licences.

Here we introduce the simplified workflow of the FirePlex-HT® immunoassays with data demonstrating the performance for quantifying key cytokines in multiplex, in biological samples. In addition, we show comparison data generated using multiple different high content imagers, which demonstrate comparable assay performance. Together, this novel combination of multiplexed, high­-sensitivity assays and bioinformatics tools enables rapid quantitation of protein biomarker signatures in biofluid specimens.

Bianca Heinrich

Senior Scientist
Abcam, Inc
Cambridge, MA

Bianca Heinrich, PhD, Senior Scientist, Abcam, Cambridge, MA 02140, USA.

I'm currently developing novel high-throughput multiplex immunoassays with Abcam in Cambridge, MA. We take advantage of our extensive repertoire of high quality antibodies to allow for higher multiplexing under no wash conditions using the FirePlex platform. Fast data analysis is realized via read-out on high-content imagers (HCI) and integrated software tools. Originally from Germany, I received my PhD in Cell and Molecular Biology from Boston University and continued my education as a postdoctoral fellow at Harvard Medical School, where I studied the life cycle of negative-strand RNA viruses.