EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes

Jacques Dongen; L Lhermitte; S Bottcher; J Almeida; Vincent van der Velden; J Flores-Montero; A Rawstron; V Asnafi; Q Lecrevisse; P Lucio; E Mejstrikova; Tomek Szczepanski; T Kalina; R de Tute; M Bruggemann; L Sedek; M Cullen; Ton Langerak; A De Mendonca; E Macintyre; M Martin-Ayuso; O Hrusak; MB Vidriales; A Orfao

doi:10.1038/leu.2012.120

EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes

Jacques Dongen, L Lhermitte, S Bottcher, J Almeida, Vincent van der Velden, J Flores-Montero, A Rawstron, V Asnafi, Q Lecrevisse, P Lucio, E Mejstrikova, Tomek Szczepanski, T Kalina, R de Tute, M Bruggemann, L Sedek, M Cullen, Ton Langerak, A De Mendonca, E MacintyreM Martin-Ayuso, O Hrusak, MB Vidriales, A Orfao

Immunology

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Abstract

Most consensus leukemia & lymphoma antibody panels consist of lists of markers based on expert opinions, but they have not been validated. Here we present the validated EuroFlow 8-color antibody panels for immunophenotyping of hematological malignancies. The single-tube screening panels and multi-tube classification panels fit into the EuroFlow diagnostic algorithm with entries defined by clinical and laboratory parameters. The panels were constructed in 2-7 sequential design-evaluation-redesign rounds, using novel Infinicyt software tools for multivariate data analysis. Two groups of markers are combined in each 8-color tube: (i) backbone markers to identify distinct cell populations in a sample, and (ii) markers for characterization of specific cell populations. In multi-tube panels, the backbone markers were optimally placed at the same fluorochrome position in every tube, to provide identical multidimensional localization of the target cell population(s). The characterization markers were positioned according to the diagnostic utility of the combined markers. Each proposed antibody combination was tested against reference databases of normal and malignant cells from healthy subjects and WHO-based disease entities, respectively. The EuroFlow studies resulted in validated and flexible 8-color antibody panels for multidimensional identification and characterization of normal and aberrant cells, optimally suited for immunophenotypic screening and classification of hematological malignancies.

Original language	Undefined/Unknown
Pages (from-to)	1908-1975
Number of pages	68
Journal	Leukemia
Volume	26
Issue number	9
DOIs	https://doi.org/10.1038/leu.2012.120
Publication status	Published - 2012

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Cite this

Dongen, J., Lhermitte, L., Bottcher, S., Almeida, J., van der Velden, V., Flores-Montero, J., Rawstron, A., Asnafi, V., Lecrevisse, Q., Lucio, P., Mejstrikova, E., Szczepanski, T., Kalina, T., de Tute, R., Bruggemann, M., Sedek, L., Cullen, M., Langerak, T., De Mendonca, A., ... Orfao, A. (2012). EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. Leukemia, 26(9), 1908-1975. https://doi.org/10.1038/leu.2012.120

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title = "EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes",

abstract = "Most consensus leukemia & lymphoma antibody panels consist of lists of markers based on expert opinions, but they have not been validated. Here we present the validated EuroFlow 8-color antibody panels for immunophenotyping of hematological malignancies. The single-tube screening panels and multi-tube classification panels fit into the EuroFlow diagnostic algorithm with entries defined by clinical and laboratory parameters. The panels were constructed in 2-7 sequential design-evaluation-redesign rounds, using novel Infinicyt software tools for multivariate data analysis. Two groups of markers are combined in each 8-color tube: (i) backbone markers to identify distinct cell populations in a sample, and (ii) markers for characterization of specific cell populations. In multi-tube panels, the backbone markers were optimally placed at the same fluorochrome position in every tube, to provide identical multidimensional localization of the target cell population(s). The characterization markers were positioned according to the diagnostic utility of the combined markers. Each proposed antibody combination was tested against reference databases of normal and malignant cells from healthy subjects and WHO-based disease entities, respectively. The EuroFlow studies resulted in validated and flexible 8-color antibody panels for multidimensional identification and characterization of normal and aberrant cells, optimally suited for immunophenotypic screening and classification of hematological malignancies.",

author = "Jacques Dongen and L Lhermitte and S Bottcher and J Almeida and {van der Velden}, Vincent and J Flores-Montero and A Rawstron and V Asnafi and Q Lecrevisse and P Lucio and E Mejstrikova and Tomek Szczepanski and T Kalina and {de Tute}, R and M Bruggemann and L Sedek and M Cullen and Ton Langerak and {De Mendonca}, A and E Macintyre and M Martin-Ayuso and O Hrusak and MB Vidriales and A Orfao",

year = "2012",

doi = "10.1038/leu.2012.120",

language = "Undefined/Unknown",

volume = "26",

pages = "1908--1975",

journal = "Leukemia",

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Dongen, J, Lhermitte, L, Bottcher, S, Almeida, J, van der Velden, V, Flores-Montero, J, Rawstron, A, Asnafi, V, Lecrevisse, Q, Lucio, P, Mejstrikova, E, Szczepanski, T, Kalina, T, de Tute, R, Bruggemann, M, Sedek, L, Cullen, M, Langerak, T, De Mendonca, A, Macintyre, E, Martin-Ayuso, M, Hrusak, O, Vidriales, MB & Orfao, A 2012, 'EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes', Leukemia, vol. 26, no. 9, pp. 1908-1975. https://doi.org/10.1038/leu.2012.120

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T1 - EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes

AU - Dongen, Jacques

AU - Lhermitte, L

AU - Bottcher, S

AU - Almeida, J

AU - van der Velden, Vincent

AU - Flores-Montero, J

AU - Rawstron, A

AU - Asnafi, V

AU - Lecrevisse, Q

AU - Lucio, P

AU - Mejstrikova, E

AU - Szczepanski, Tomek

AU - Kalina, T

AU - de Tute, R

AU - Bruggemann, M

AU - Sedek, L

AU - Cullen, M

AU - Langerak, Ton

AU - De Mendonca, A

AU - Macintyre, E

AU - Martin-Ayuso, M

AU - Hrusak, O

AU - Vidriales, MB

AU - Orfao, A

PY - 2012

Y1 - 2012

N2 - Most consensus leukemia & lymphoma antibody panels consist of lists of markers based on expert opinions, but they have not been validated. Here we present the validated EuroFlow 8-color antibody panels for immunophenotyping of hematological malignancies. The single-tube screening panels and multi-tube classification panels fit into the EuroFlow diagnostic algorithm with entries defined by clinical and laboratory parameters. The panels were constructed in 2-7 sequential design-evaluation-redesign rounds, using novel Infinicyt software tools for multivariate data analysis. Two groups of markers are combined in each 8-color tube: (i) backbone markers to identify distinct cell populations in a sample, and (ii) markers for characterization of specific cell populations. In multi-tube panels, the backbone markers were optimally placed at the same fluorochrome position in every tube, to provide identical multidimensional localization of the target cell population(s). The characterization markers were positioned according to the diagnostic utility of the combined markers. Each proposed antibody combination was tested against reference databases of normal and malignant cells from healthy subjects and WHO-based disease entities, respectively. The EuroFlow studies resulted in validated and flexible 8-color antibody panels for multidimensional identification and characterization of normal and aberrant cells, optimally suited for immunophenotypic screening and classification of hematological malignancies.

AB - Most consensus leukemia & lymphoma antibody panels consist of lists of markers based on expert opinions, but they have not been validated. Here we present the validated EuroFlow 8-color antibody panels for immunophenotyping of hematological malignancies. The single-tube screening panels and multi-tube classification panels fit into the EuroFlow diagnostic algorithm with entries defined by clinical and laboratory parameters. The panels were constructed in 2-7 sequential design-evaluation-redesign rounds, using novel Infinicyt software tools for multivariate data analysis. Two groups of markers are combined in each 8-color tube: (i) backbone markers to identify distinct cell populations in a sample, and (ii) markers for characterization of specific cell populations. In multi-tube panels, the backbone markers were optimally placed at the same fluorochrome position in every tube, to provide identical multidimensional localization of the target cell population(s). The characterization markers were positioned according to the diagnostic utility of the combined markers. Each proposed antibody combination was tested against reference databases of normal and malignant cells from healthy subjects and WHO-based disease entities, respectively. The EuroFlow studies resulted in validated and flexible 8-color antibody panels for multidimensional identification and characterization of normal and aberrant cells, optimally suited for immunophenotypic screening and classification of hematological malignancies.

U2 - 10.1038/leu.2012.120

DO - 10.1038/leu.2012.120

M3 - Article

SN - 0887-6924

VL - 26

SP - 1908

EP - 1975

JO - Leukemia

JF - Leukemia

IS - 9

ER -