Analysis of individual data from bead-based assays ("bead arrays")

James W. Jacobson, Kerry G. Oliver, Christy Weiss, John Kettman

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

Introduction: Typically, bead-based assays ("bead arrays") use the mean or median value of a population of measurements to judge ligand binding or other activity, which results in a change in fluorescence intensity. Individual bead measurements are used here to calculate population parameters integral to the measurement of a bead array. Methods: Using a commercially-available instrument designed for bead array measurements, a set of beads were labeled with biotin and then titrated with PE-Streptavidin. Data were collected under normal machine conditions as well as variations. Results: The "sensitivity" of the measurements was determined using parametric and nonparametric statistical methods as well as regression analysis over a limited range of the titration (concentration vs. response profile). Conclusions: Results at low ligand concentrations suggest that precise measurements with bead array systems require a large number of individual bead measurements to be acquired. Individual bead measurements should be used to determine the mean and confidence intervals for the calculated measurements. These results also apply to regression analysis of concentration-response profiles. Furthermore, features of the instrument can be manipulated to achieve increased sensitivity and detection of lower amounts of ligand bound to the bead populations.

Original languageEnglish (US)
Pages (from-to)384-390
Number of pages7
JournalCytometry Part A
Volume69
Issue number5
DOIs
Publication statusPublished - May 2006

    Fingerprint

Keywords

  • Bead arrays
  • Biotin-avidin
  • Coefficient of variation
  • Confidence intervals
  • Luminex 100
  • Sensitivity of bead measurements
  • Statistical testing

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Jacobson, J. W., Oliver, K. G., Weiss, C., & Kettman, J. (2006). Analysis of individual data from bead-based assays ("bead arrays"). Cytometry Part A, 69(5), 384-390. https://doi.org/10.1002/cyto.a.20293