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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
Insights
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.
Abstract:
As quantitative flow cytometry is being increasingly used to characterize non-malignant and malignant disorders, interlaboratory standardization becomes an important issue. However, the lack of standardized methods and process controls with predefined antibody binding capacity values, limits direct interlaboratory comparison. The present study has addressed these issues using a stable whole blood product and a standardized antigen quantification protocol. It was demonstrated that: (i) a standard technical protocol can result in a high degree of interlaboratory concordance; (ii) interlaboratory variation of less than 12% can be achieved for CD4 antibody binding capacity values; and (iii) stable whole blood can be used as a process control with predefined antibody binding capacity values. Furthermore, using such an approach, a normal range was established for CD3, CD4 CD8 and CD19. These antigens appear to be expressed in a hierarchical manner, a factor that could be used as a procedural quality control measure.
