Comparison of poly- and monoclonal antibodies for determination of B-cell clonal excess in an routine clinical laboratory.
Flow cytometry (FCM) has gained wide use in the determination of clonality in B-cell lymphoproliferative diseases and many methodological variations exist. We have compared the suitability of a) dual fluorochrome (FITC/PE)-labelled monoclonal antibodies, b) single fluorochrome (FITC)-labelled monoclonal antibodies and c) F(ab')2 fragments of FITC-labelled polyclonal antibodies for flow cytometric determination of clonality using commercially available software and a short sample preparation protocol. The FCM method was validated by analysis of immunoglobulin heavy chain and light chain gene rearrangements. We recommend the use of FITC-labelled monoclonals to obtain three parameters, the kappa/lambda ratio, D and D/S(n) values (Kolmogorov-Smirnov statistics) instead of the commonly used kappa/lambda ratio and D values only. This allows the use of a rapid sample preparation protocol to blood and bone marrow aspirates without sacrificing sensitivity or specificity obtained by the usual FCM method.
Agrawal
Y P
YP
Department of Clinical Chemistry, Kuopio University Hospital, Finland.
Hämäläinen
E
E
Mahlamäki
E K
EK
Aho
H
H
Nousianinen
T
T
Lahtinen
R
R
Penttilä
I M
IM
D000906:Antibodies; D000911:Antibodies, Monoclonal; D001402:B-Lymphocytes / Q000276:immunology*; D001853:Bone Marrow / Q000276:immunology / Q000473:pathology; D005434:Flow Cytometry / Q000379:methods; D016650:Fluorescein-5-isothiocyanate; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D016130:Immunophenotyping; D007938:Leukemia / Q000276:immunology*; D008223:Lymphoma / Q000276:immunology*; D008232:Lymphoproliferative Disorders / Q000276:immunology* / Q000473:pathology
D000906:Antibodies; D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D016650:Fluorescein-5-isothiocyanate