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Related Experiment Videos

Performance evaluation of QuantiBRITE phycoerythrin beads.

K K Pannu1, E T Joe, S B Iyer

  • 1BD Biosciences, San Jose, California 95131, USA.

Cytometry
|December 18, 2001
PubMed
Summary

QuantiBRITE phycoerythrin (PE) beads offer a reliable method for standardizing antibody bound per cell (ABC) quantitation. This tool ensures consistent results across different laboratories and over time for CD4 and CD69 analysis.

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Area of Science:

  • Immunology
  • Flow Cytometry
  • Biotechnology

Background:

  • Standardization of antibody quantitation is crucial for reliable flow cytometry analysis.
  • Phycoerythrin (PE) beads offer a potential solution for standardizing measurements of antibodies bound per cell (ABC).

Purpose of the Study:

  • To evaluate the performance of QuantiBRITE PE beads for standardizing CD4 and CD69 quantitation.
  • To assess the precision, inter-instrument variability, and temporal stability of PE bead-based standardization.

Main Methods:

  • Whole blood was stained for CD4 quantitation using a no-wash/no-lyse format.
  • Whole blood was activated and stained for CD69 quantitation using a lyse-no-wash format.
  • QuantiBRITE PE beads were used to standardize antibody bound per cell (ABC) measurements.

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Main Results:

  • CD4 ABC quantitation showed high precision (<1.0% CV within-assay, <5.0% CV across instruments).
  • CD4 ABC quantitation demonstrated good temporal stability (1.3%-3.2% CV within-donor over 2 months).
  • CD69 ABC quantitation showed acceptable precision (<3.0% CV within-assay) but higher temporal variability (2%-24% CV within-donor over 1 month).

Conclusions:

  • QuantiBRITE PE beads are a valuable tool for standardized flow cytometry analysis across laboratories.
  • The beads provide a simple and robust method for quantitating antibody expression levels in terms of ABC when used with 1:1 PE conjugates.