title: Comparison of poly- and monoclonal antibodies for determination of B-cell clonal excess in an routine clinical laboratory.
abstract: 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.
authors.affiliation: Department of Clinical Chemistry, Kuopio University Hospital, Finland.
authors.forename: Y P
authors.initials: YP
authors.lastname: Agrawal
authors.forename: E
authors.initials: E
authors.lastname: Hämäläinen
authors.forename: E K
authors.initials: EK
authors.lastname: Mahlamäki
authors.forename: H
authors.initials: H
authors.lastname: Aho
authors.forename: T
authors.initials: T
authors.lastname: Nousianinen
authors.forename: R
authors.initials: R
authors.lastname: Lahtinen
authors.forename: I M
authors.initials: IM
authors.lastname: Penttilä
mesh_terms: 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
chemical_list: D000906:Antibodies; D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D016650:Fluorescein-5-isothiocyanate