Protocol

Serum/Plasma Proteomics

Volume 728 of the series Methods in Molecular Biology pp 195-206

Date:

A Fluorescent Microsphere-Based Method for Assay of Multiple Analytes in Plasma

  • Oliver K. Bernhard
  • , Rommel A. Mathias
  • , Thomas W. Barnes
  • , Richard J. SimpsonAffiliated withLudwig Institute for Cancer Research, Royal Melbourne Hospital Email author 

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Abstract

Measurement of multiple analytes can provide increased sensitivity and specificity for the detection and management of disease. The enzyme-linked immunosorbent assay (ELISA) is currently the “gold standard” for protein quantification; however, individual assays for each analyte must be performed, placing demand on sample volume. On the contrary, multiplex assays using microsphere-based technologies allow for multiple analytes to be simultaneously assayed within a single sample. Here, we present a protocol for the preparation and development of a multiple-analyte assay in human plasma using the BioPlex 200 platform (Bio-Rad), which incorporates xMAP technology (Luminex).

Key words

Fluorescent beads Multiplex assay Biomarker validation Flow cytometry Plasma