Bead-Based Assays for Validating Proteomic Profiles in Body Fluids

Methods Mol Biol. 2021;2344:65-78. doi: 10.1007/978-1-0716-1562-1_5.

Abstract

Protein biomarkers in biological fluids represent an important resource for improving the clinical management of diseases. Current proteomics technologies are capable of performing high-throughput and multiplex profiling in different types of fluids, often leading to the shortlisting of tens of candidate biomarkers per study. However, before reaching any clinical setting, these discoveries require thorough validation and an assay that would be suitable for routine analyses. In the path from biomarker discovery to validation, the performance of the assay implemented for the intended protein quantification is extremely critical toward achieving reliable and reproducible results. Development of robust sandwich immunoassays for individual candidates is challenging and labor and resource intensive, and multiplies when evaluating a panel of interesting candidates at the same time. Here we describe a versatile pipeline that facilitates the systematic and parallel development of multiple sandwich immunoassays using a bead-based technology.

Keywords: Antibody; Mass spectrometry (MS); Protein standard; Sandwich immunoassay (SIA); Single-binder immunoassays; Suspension bead array (SBA).

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Biomarkers / analysis
  • Body Fluids / chemistry*
  • Enzyme-Linked Immunosorbent Assay*
  • Humans
  • Proteome / analysis*
  • Proteomics*

Substances

  • Biomarkers
  • Proteome