Immunomagnetic isolation of magnetoferritin-labeled cells in a modified ferrograph.
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 field 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 field of 1.67 Tesla (T), a field gradient of 2.57 T/mm, and a flow 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.
Zborowski
M
M
Department of Biomedical Engineering/Wb-3, Cleveland Clinic Foundation, OH 44195-5254, USA. zborow@bme.ri.ccf.org
Fuh
C B
CB
Green
R
R
Baldwin
N J
NJ
Reddy
S
S
Douglas
T
T
Mann
S
S
Chalmers
J J
JJ
D000818:Animals; D001052:Apoferritins / Q000031:analogs & derivatives* / Q000737:chemistry; D017252:CD3 Complex / Q000032:analysis*; D015704:CD4 Antigens / Q000032:analysis*; D016827:CD8 Antigens / Q000032:analysis*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D006801:Humans; D018189:Immunomagnetic Separation; D007501:Iron / Q000737:chemistry*; D051379:Mice; D010087:Oxides / Q000737:chemistry*
D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens; D010087:Oxides; C088922:magnetoferritin; D001052:Apoferritins; D007501:Iron; D016650:Fluorescein-5-isothiocyanate