Fast and Efficient Measurement of Clinical and Biological Samples Using Immunoassay-Based Multiplexing Systems

Methods Mol Biol. 2020:2102:129-147. doi: 10.1007/978-1-0716-0223-2_6.

Abstract

Immunoassay is one of the most commonly used biomedical techniques to detect the expression of an antibody or an antigen in a test sample. Enzyme-linked immunosorbent assay (ELISA) has been used for a variety of applications including diagnostic tools and quality controls. However, one of the main limitations of ELISA is its lack of multiplexing ability, so ELISA may not be an efficient diagnostic tool when a measurement of multiple determinants is needed for samples with limited quantity such as blood or biological samples from newborns or babies. Although similar to ELISA in assay measurement, multiplex platforms such as bead-based Luminex and multi-array-based MSD (Meso Scale Discovery) are widely used to measure multiple biomarkers from a single analysis. Luminex is a xMAP-based technology that combines several different technologies to provide an efficient and accurate measurement of multiple analytes from a single sample. The multiplexing can be achieved because up to 100 distinct Luminex color-coded microsphere bead sets can be coated with a reagent specific to a particular bioassay, allowing the capture and detection of specific analytes from a sample. Using Multi-array and electrochemiluminescence technologies, the MSD platform provides the multiplex capability with similar consistence as observed in ELISA. Various biological samples that can be analyzed by both Luminex and MSD systems include serum, plasma, tissue and cell lysate, saliva, sputum, and bronchoalveolar Lavage (BAL). The most common Luminex and MSD-based assays are to detect a combined set of cytokines to provide a measurement of cytokine expression profiling for a diagnostic purpose.

Keywords: Biofluid; Biomarker; Luminex; MSD; Multiplex.

MeSH terms

  • Animals
  • Antibodies / immunology*
  • Biomarkers / blood*
  • Bronchoalveolar Lavage Fluid / immunology
  • Cytokines / blood
  • Enzyme-Linked Immunosorbent Assay / methods
  • Fluorescence
  • Humans
  • Immunoassay / instrumentation*
  • Immunoassay / methods*
  • Lung / immunology
  • Lung / metabolism
  • Mice
  • Plasma / immunology
  • Plasma / metabolism
  • Reagent Kits, Diagnostic
  • Serum / immunology
  • Serum / metabolism
  • Workflow

Substances

  • Antibodies
  • Biomarkers
  • Cytokines
  • Reagent Kits, Diagnostic