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Quantitation of CD38 expression using QuantiBRITE beads
S B Iyer1, L E Hultin, J A Zawadzki
1Becton Dickinson Immunocytometry Systems, San Jose, California, USA. sujata_iyer@bdis.com
Cytometry
|October 17, 1998
Summary
The QuantiBRITE bead method reliably quantifies CD38 expression on CD8+ T-lymphocytes. This method, using specific repurified antibodies, aligns with the CD4 biological calibration method for accurate results in HIV research.
Area of Science:
- Immunology
- Virology
- Biotechnology
Background:
- CD38 is a marker on CD8+ T-lymphocytes, crucial for understanding HIV pathogenesis.
- Accurate quantitation of CD38 expression is essential for prognostic interpretation.
Purpose of the Study:
- To compare the QuantiBRITE bead method with the CD4 biological calibration method for quantifying CD38 expression.
- To establish conditions for consistent and reliable CD38 quantitation.
Main Methods:
- Comparison of QuantiBRITE and CD4 biological calibration methods.
- Use of repurified CD38-phycoerythrin (PE) and CD4-PE conjugates.
- Analysis of CD38 expression on CD8+ T-lymphocytes from Multicenter AIDS Cohort Study participants.
Main Results:
- The QuantiBRITE method yielded identical results to the CD4 biological calibration method under specific conditions.
- Repurified CD38-PE and CD4-PE are critical for accurate quantitation.
- Standardized sample preparation methods (NH4Cl, FACS lysing, buffer dilution, paraformaldehyde fixation) showed consistent CD38 relative fluorescence intensity.
Conclusions:
- Repurified PE conjugates of CD38 and QuantiBRITE PE beads offer a convenient and reliable method for CD38 quantitation.
- Prognostic significance of QuantiBRITE-derived CD38 values can be interpreted using established reports.
- Understanding antibody valence is important for antigen density determination from antibodies bound per cell (ABCs).