Immunomagnetic isolation of magnetoferritin-labeled cells in a modified ferrograph

Maciej Zborowski, Chwan Bor Fuh, Ralph Green, Nicholas J. Baldwin, Sridhar Reddy, Trevor Douglas, Stephen Mann, Jeffrey J. Chalmers

Research output: Contribution to journalArticle

39 Scopus citations

Abstract

Pan T, helper, and cytotoxic lymphocytes were isolated from the human peripheral blood mononuclear cell fraction by antibody staining, ferritin labeling, and deposition on glass slides. Two distinct forms of ferritin were used: one was native horse spleen ferritin, and the other was magnetoferritin. Magnetoferritin was obtained by reconstituting the horse spleen ferritin iron core with superparamagnetic magnetite instead of the usual paramagnetic ferrihydrite crystal. The cell deposition on microscopic glass slides in the magnetic held was obtained by an instrument that was adapted from an industrial magnetic deposition analyzer, the ferrograph. The identity of cells in the magnetic deposits was confirmed by comparing the cell fractions in the feed and in the eluate with the use of flow cytometry. The immunostaining protocol amplified the number of ferritin molecules per cell surface antigen 20-70 times. Magnetoferritin, but not native ferritin, imparted a sufficient magnetic moment to cells to deplete the labeled cell population between 67 and 88% of its initial concentration in a magnetic held of 1.67 Tesla (T), a field gradient of 2.57 T/mm, and a now rate of 0.01 ml/min. This study showed that the magnetic moment of magnetoferritin was sufficient for immunomagnetic isolation of lymphocytes from mononuclear cell preparations in the modified ferrograph.

Original languageEnglish (US)
Pages (from-to)251-259
Number of pages9
JournalCytometry
Volume24
Issue number3
DOIs
StatePublished - Jul 1 1996
Externally publishedYes

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Keywords

  • CD3
  • CD4
  • CD8
  • Cytotoxic T cell
  • Ferritin
  • Helper T cell
  • Immunomagnetic separation
  • Lymphocyte immunophenotyping
  • Magnetoferritin
  • Pan T cell

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Zborowski, M., Fuh, C. B., Green, R., Baldwin, N. J., Reddy, S., Douglas, T., Mann, S., & Chalmers, J. J. (1996). Immunomagnetic isolation of magnetoferritin-labeled cells in a modified ferrograph. Cytometry, 24(3), 251-259. https://doi.org/10.1002/(SICI)1097-0320(19960701)24:3<251::AID-CYTO8>3.0.CO;2-K