Standardization of lymphocyte antibody binding capacity - a multi-centre study.
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.
Barnett
D
D
UK NEQAS for Leucocyte Immunophenotyping, Royal Hallamshire Hospital (co-ordinating centre), Sheffield, UK. d.barnett@sheffield.ac.uk
Storie
I
I
Granger
V
V
Whitby
L
L
Reilly
J T
JT
Brough
S
S
Garner
S
S
Lawry
J
J
Richards
S
S
Bell
A E
AE
Shenton
B K
BK
D015703:Antigens, CD / Q000097:blood / Q000276:immunology; D000954:Antigens, Surface / Q000032:analysis*; D001666:Binding Sites, Antibody; D002985:Clinical Protocols / Q000592:standards; D005260:Female; D005434:Flow Cytometry / Q000592:standards*; D006801:Humans; D007150:Immunohistochemistry / Q000592:standards; D007519:Isoantigens / Q000097:blood / Q000276:immunology; D008214:Lymphocytes / Q000097:blood / Q000737:chemistry / Q000276:immunology*; D008297:Male; D015588:Observer Variation; D011786:Quality Control; D012015:Reference Standards; D012737:Sex Factors; D013997:Time Factors
D015703:Antigens, CD; D000954:Antigens, Surface; D007519:Isoantigens