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Characterization of antibody binding to three cancer-related antigens using flow cytometry and cell tracking velocimetry
Biotechnology and bioengineering, 2003-05, Vol.82 (3), p.340-351
Chosy, E. Julia
Nakamura, Masayuki
Melnik, Kristie
Comella, Kristin
Lasky, Larry C.
Zborowski, Maciej
Chalmers, Jeffrey J.
2003
Volltextzugriff (PDF)
Details
Autor(en) / Beteiligte
Chosy, E. Julia
Nakamura, Masayuki
Melnik, Kristie
Comella, Kristin
Lasky, Larry C.
Zborowski, Maciej
Chalmers, Jeffrey J.
Titel
Characterization of antibody binding to three cancer-related antigens using flow cytometry and cell tracking velocimetry
Ist Teil von
Biotechnology and bioengineering, 2003-05, Vol.82 (3), p.340-351
Ort / Verlag
New York: Wiley Subscription Services, Inc., A Wiley Company
Erscheinungsjahr
2003
Quelle
MEDLINE
Beschreibungen/Notizen
Proper antibody labeling is a fundamental step in the positive selection/isolation of rare cancer cells using immunomagnetic cell separation technology. Using either a two‐step or single‐step labeling protocol, we examined a combination of six different antibodies specific for three different antigens (epithelial specific antigen, epithelial membrane antigen, and HER‐2/Neu) on two different breast cancer cell lines (HCC1954 and MCF‐7). When a two‐step labeling protocol was used (i.e., anti‐surface marker–fluoroscein‐isothiocyanate [FITC] [primary Ab], anti‐FITC magnetic colloid [secondary Ab]) saturation of the primary antibody was determined using fluorescence intensity measurements from flow cytometry (FCM). The saturation of the secondary antibody (or saturation of a single‐step labeling) was determined using magnetophoretic mobility measurements from cell tracking velocimetry (CTV). When the maximum magnetophoretic mobility was the primary objective, our results demonstrate that the quantities necessary for antibody saturation with respect to fluorescence intensity were generally higher than those recommended by the manufacturer. The results demonstrate that magnetophoretic mobility varies depending on the types of cell lines, primary antibodies, and concentration of secondary magnetic colloid‐conjugated antibody. It is concluded that saturation studies are a vital preparatory step in any separation method involving antibody labeling, especially those that require the specificity of rare cell detection. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 82: 340–351, 2003.
Sprache
Englisch
Identifikatoren
ISSN: 0006-3592
eISSN: 1097-0290
DOI: 10.1002/bit.10581
Titel-ID: cdi_proquest_miscellaneous_73034073
Format
–
Schlagworte
Antigen-Antibody Reactions - immunology
,
Antigens, Neoplasm - immunology
,
Biological and medical sciences
,
Biomarkers, Tumor - immunology
,
Biotechnology
,
breast carcinoma
,
Breast Neoplasms - classification
,
Breast Neoplasms - immunology
,
Breast Neoplasms - metabolism
,
Breast Neoplasms - pathology
,
CA 15-3
,
CA 17-1
,
Cell Movement
,
Cell Separation - methods
,
cell tracking velocimetry
,
flow cytometry
,
Flow Cytometry - methods
,
Fundamental and applied biological sciences. Psychology
,
Health. Pharmaceutical industry
,
HER-2/Neu
,
Humans
,
immunomagnetic cytochemistry
,
Immunomagnetic Separation - methods
,
Industrial applications and implications. Economical aspects
,
magnetic cell separation
,
Miscellaneous
,
Rheology - methods
,
Tumor Cells, Cultured
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