Immunomagnetic enrichment of CD138 positive cells from weakly infiltrated myeloma patients samples enables the determination of the tumor clone specific IgH rearrangement.
In multiple myeloma, the polymerase chain reaction (PCR) of the Ig heavy chain with allele-specific oligonucleotide (ASO) primers is a common and well-described method of identifying the tumor clone in peripheral blood (PB), bone marrow (BM) or leukapheresis products (LA). A factor which is crucial to the detection of clonal Ig rearrangements lies in the 'purity' of the tumor tissue used for the consensus PCR. We describe the application of a method to enrich CD138 positive myeloma cells derived from weakly infiltrated PB-, BM- and LA-samples. These are subjected to immunomagnetic enrichment with the MACS system, using an CD138 antibody directly conjugated to magnetic beads to obtain an enriched tumor cell population and the subsequent amplification of tumor specific IgH rearrangements. We investigated 29 samples (ten PB, ten BM, nine LA) with a median myeloma cell content of 0.5%. The approach led to a median enrichment factor of 118. Tumor-specific rearrangements could be amplified reproducibly from samples containing less than 0.1% myeloma cells.
Department of Internal Medicine I, University of Cologne, 50924 Cologne, Germany. Andreas.Draube@uni-koeln.de
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Draube
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Pfister
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Vockerodt
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Schuster
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Kube
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Diehl
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Tesch
D016384:Consensus Sequence; D015328:Gene Rearrangement, B-Lymphocyte, Light Chain; D006801:Humans; D018189:Immunomagnetic Separation; D008562:Membrane Glycoproteins / Q000032:analysis*; D009101:Multiple Myeloma / Q000276:immunology* / Q000473:pathology*; D016133:Polymerase Chain Reaction; D011509:Proteoglycans / Q000032:analysis*; D017421:Sequence Analysis; D053668:Syndecan-1; D053667:Syndecans
D008562:Membrane Glycoproteins; D011509:Proteoglycans; C508241:SDC1 protein, human; D053668:Syndecan-1; D053667:Syndecans