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Standardization of lymphocyte antibody binding capacity - a multi-centre study.
D Barnett1, I Storie, V Granger
1UK NEQAS for Leucocyte Immunophenotyping, Royal Hallamshire Hospital (co-ordinating centre), Sheffield, UK. d.barnett@sheffield.ac.uk
Clinical and Laboratory Haematology
|May 3, 2000
Summary
Standardizing quantitative flow cytometry is crucial for comparing lab results. This study shows a stable whole blood product and protocol achieve high interlaboratory concordance for antigen quantification, enabling reliable normal range establishment.
Area of Science:
- Immunology
- Clinical Laboratory Science
- Biotechnology
Background:
- Quantitative flow cytometry is essential for diagnosing various disorders, but interlaboratory standardization remains a challenge.
- Lack of standardized methods and process controls hinders direct comparison of flow cytometry data between laboratories.
- Predefined antibody binding capacity values are needed for robust standardization.
Purpose of the Study:
- To address interlaboratory standardization issues in quantitative flow cytometry.
- To evaluate the use of a stable whole blood product as a process control.
- To establish a standardized antigen quantification protocol for improved laboratory concordance.
Main Methods:
- A stable whole blood product was utilized for process control.
- A standardized antigen quantification protocol was developed and applied.
- Interlaboratory comparisons were performed to assess concordance and variation.
Main Results:
- A standard technical protocol achieved high interlaboratory concordance.
- Interlaboratory variation for CD4 antibody binding capacity was less than 12%.
- Stable whole blood proved effective as a process control with predefined values.
Conclusions:
- Standardized protocols and stable whole blood controls enhance interlaboratory comparability in flow cytometry.
- This approach enabled the establishment of normal ranges for CD3, CD4, CD8, and CD19.
- Antigen expression hierarchy can serve as a valuable procedural quality control measure.