Document Detail

Rapid enrichment and detection of melanoma cells from peripheral blood mononuclear cells by a new assay combining immunomagnetic cell sorting and immunocytochemical staining.
MedLine Citation:
PMID:  11092033     Owner:  NLM     Status:  MEDLINE    
Commonly used methods for detection of melanoma cells in blood, including RT-PCR and immunocytochemistry, display only a limited sensitivity and specificity. Reliable detection of less than one melanoma cell per ml of blood is hardly possible using these methods. To obtain greater sensitivity so that a single melanoma cell in up to 25 ml of blood can be detected (5 x 10(7) peripheral blood mononuclear cells, or PBMC), we developed a new assay for combined enrichment and immunocytochemical detection of disseminated melanoma cells from PBMC of patients with malignant melanomas. Melanoma cells are directly magnetically labeled using colloidal superparamagnetic microparticles approximately 60 nm in diameter conjugated to the anti-melanoma monoclonal antibody 9.2.27, with no reactivity to normal cells in blood. Magnetically labeled melanoma cells are enriched from PBMC by magnetic cell separation and detected by a new approach for immunocytochemical staining with monoclonal mouse anti-melanoma antibodies (anti-MelanA and HMB-45). The efficiency of this assay was demonstrated in a model system in which 5-500 tumor cells from the melanoma cell line SK-MEL-28 were seeded into PBMC samples from healthy donors containing 5 x 10(7) leukocytes. Mean recovery of the seeded tumor cells was 47.4 +/- 13.99% (n = 15). Applying the assay to 20-50 ml blood samples of patients with stage III-IV malignant melanomas, we were able to detect melanoma cells in two of eight patients (25%).
C Siewert; M Herber; N Hunzelmann; O Fodstad; S Miltenyi; M Assenmacher; J Schmitz
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Publication Detail:
Type:  Comparative Study; Journal Article    
Journal Detail:
Title:  Recent results in cancer research. Fortschritte der Krebsforschung. Progrès dans les recherches sur le cancer     Volume:  158     ISSN:  0080-0015     ISO Abbreviation:  Recent Results Cancer Res.     Publication Date:  2001  
Date Detail:
Created Date:  2001-02-14     Completed Date:  2001-03-15     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  0044671     Medline TA:  Recent Results Cancer Res     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  51-60     Citation Subset:  IM    
Miltenyi Biotec GmbH, Bergisch Gladbach, Germany.
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MeSH Terms
Antibodies, Monoclonal
Antigens, Neoplasm
Immunohistochemistry / methods*
Immunomagnetic Separation / methods*
Leukocytes, Mononuclear / pathology*
Melanoma / diagnosis*
Neoplasm Metastasis / diagnosis
Neoplasm Proteins / analysis
Neoplasm Staging
Neoplastic Cells, Circulating / pathology*
Tumor Cells, Cultured
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Antigens, Neoplasm; 0/MART1 antigen; 0/Neoplasm Proteins

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