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2 results on '"Koning-Goedheer A"'

1. Detection of fusion genes at the protein level in leukemia patients via the flow cytometric immunobead assay

Author

Tomas Kalina, V H J van der Velden, F J T Staal, P G Hoogeveen, Alberto Orfao, Paulo Sérgio Lucio, Monika Brüggemann, Martin Kovac, Nancy Boeckx, Edwin Sonneveld, Juan Flores-Montero, A. Koning-Goedheer, Lukasz Sedek, W.M. Comans-Bitter, J. J. M. Van Dongen, Paul Evans, E Dekking, S Böttcher, Tomasz Szczepański, Immunology, and Molecular Genetics

Subjects
Immunoassay, Acute leukemia, Leukemia, Oncogene Proteins, Oncogene Proteins, Fusion, Clinical Biochemistry, Biology, Molecular diagnostics, Flow Cytometry, Chromosome aberration, Fusion protein, Antibodies, Immunophenotyping, Fusion gene, Oncology, hemic and lymphatic diseases, Cancer research, Humans, Oncogene Fusion, Pathology, Molecular
Abstract

Nowadays, the presence of specific genetic aberrations is progressively used for classification and treatment stratification, because acute leukemias with the same oncogenetic aberration generally form a clinically and diagnostically homogenous disease entity with comparable prognosis. Many oncogenetic aberrations in acute leukemias result in a fusion gene, which is transcribed into fusion transcripts and translated into fusion proteins, which are assumed to play a critical role in the oncogenetic process. Fusion gene aberrations are detected by karyotyping, FISH, or RT-PCR analysis. However, these molecular genetic techniques are laborious and time consuming, which is in contrast to flow cytometric techniques. Therefore we developed a flow cytometric immunobead assay for detection of fusion proteins in lysates of leukemia cell samples by use of a bead-bound catching antibody against one side of the fusion protein and fluorochrome-conjugated detection antibody. So far, we have been able to design such fusion protein immunobead assays for BCR-ABL, PML-RARA, TEL-AML1, E2A-PBX1, MLL-AF4, AML1-ETO and CBFB-MYH11. The immunobead assay for detection of fusion proteins can be performed within 3 to 4 hours in a routine diagnostic setting, without the need of special equipment other than a flow cytometer. The novel immunobead assay will enable fast and easy classification of acute leukemia patients that express fusion proteins. Such patients can be included at an early stage in the right treatment protocols, much faster than by use of current molecular techniques. The immunobead assay can be run in parallel to routine immunophenotyping and is particularly attractive for clinical settings without direct access to molecular diagnostics. © 2010 Elsevier Ltd. All rights reserved., The research and development of the flow cytometric immunobead assay and the EuroFlow Consortium were supported by the European Commission (grant STREP EU-FP6, LSHB-CT-2006- 018708).

Published
2010

2. Detection of fusion genes at the protein level in leukemia patients via the flow cytometric immunobead assay.

Author

Dekking, E., van der Velden, V.H.J., Böttcher, S., Brüggemann, M., Sonneveld, E., Koning-Goedheer, A., Boeckx, N., Lucio, P., Sedek, L., Szczepański, T., Kalina, T., Kovac, M., Evans, P., Hoogeveen, P.G., Flores-Montero, J., Orfao, A., Comans-Bitter, W.M., Staal, F.J.T., and van Dongen, J.J.M.

Subjects
LEUKEMIA, CANCER cells, ONCOGENES, GENE fusion, CYTOMETRY, MOLECULAR genetics, GENE expression, PATIENTS
Abstract

Nowadays, the presence of specific genetic aberrations is progressively used for classification and treatment stratification, because acute leukemias with the same oncogenetic aberration generally form a clinically and diagnostically homogenous disease entity with comparable prognosis. Many oncogenetic aberrations in acute leukemias result in a fusion gene, which is transcribed into fusion transcripts and translated into fusion proteins, which are assumed to play a critical role in the oncogenetic process. Fusion gene aberrations are detected by karyotyping, FISH, or RT-PCR analysis. However, these molecular genetic techniques are laborious and time consuming, which is in contrast to flow cytometric techniques. Therefore we developed a flow cytometric immunobead assay for detection of fusion proteins in lysates of leukemia cell samples by use of a bead-bound catching antibody against one side of the fusion protein and fluorochrome-conjugated detection antibody. So far, we have been able to design such fusion protein immunobead assays for BCR-ABL, PML-RARA, TEL-AML1, E2A-PBX1, MLL-AF4, AML1-ETO and CBFB-MYH11. The immunobead assay for detection of fusion proteins can be performed within 3 to 4 hours in a routine diagnostic setting, without the need of special equipment other than a flow cytometer. The novel immunobead assay will enable fast and easy classification of acute leukemia patients that express fusion proteins. Such patients can be included at an early stage in the right treatment protocols, much faster than by use of current molecular techniques. The immunobead assay can be run in parallel to routine immunophenotyping and is particularly attractive for clinical settings without direct access to molecular diagnostics. [ABSTRACT FROM AUTHOR]

Published
2010
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