Visual Interpretation Using Multiplex Immunoassay of Serum Samples

Clinical proteomic research aims to identify biomarkers that may be useful in disease diagnosis, prognosis estimation, and treatment monitoring. An immunoassay that can test multiple analytes concurrently is known as a multiplex assay. The Enzyme-linked immunosorbent assay (ELISA) format is the foundation from which multiplex immunoassays were developed and generates several signal readings as opposed to a single signal measurement. This chapter aims to provide insight into serum sample analysis interpretation based on the data output from the multiplex immunoassay platform. Although the research arena encompasses a variety of biological samples for testing, biological fluid is the matrix of choice for most multiplex immunoassays. Analysis on the immunoassay platform may allow multiplexing up to 100 different assays within a single sample. Fluorescent signals from each magnetic bead-based technology are simultaneously recorded by high-speed digital signal processors and software, which convert the signals into data. Tools for biomarker discovery-driven data analysis offer a variety of spectrum visualization options. Univariate analysis, receiver operating characteristic curves, and dot plots are used to examine differentially expressed proteins. Standard curves are automatically generated and easily optimized by the user upon visual inspection. Graphical representation of the analyte concentration levels present in the serum samples provides an easy avenue for the researcher to convey their findings. These results can be further supported with analysis using statistical software packages. Multiplex immunoassays provide several major benefits, including screening biomarkers for further assessment and confirming data for drug development. Conservation of restricted sample volume and reduction in cost per data point. Furthermore, this platform ensures increased accuracy due to reduced sample handling. This chapter further discusses the significance of selecting the best-suited commercially available kits based on optimal methods and the design of researcher-specific needs. Including taking into account present and future advancements for the multiplex immunoassay platform.