Immunomagnetic removal of B-lymphoma cells using a novel mono-sized magnetizable polymer bead, M-280, in conjunction with primary IgM and IgG antibodies.
The efficacy of a novel monosized and magnetizable polymer bead, denoted M-280, in immunomagnetic removal of Rael B-lymphoma cells was compared with that of M-450 beads, previously used in bone marrow purging. The M-280 beads which are smaller (diameter 2.8 microns) and contain less iron than the M-450 beads were coated with polyclonal IgG sheep antimouse (SAM) antibody. The two types of immunobeads were equally efficacious when used together with the mouse monoclonal IgG antibodies HH1 or FN1, giving tumor cell depletions of about 3 logs in one cycle of operation. However, when used together with the primary IgM monoclonal antibodies (MoAbs) AB1 or HH2, the new immunobeads were significantly more efficacious than the M-450 immunobeads. To elucidate the underlying mechanism flow cytometric studies and measurements of the binding of the labeled primary MoAbs to the cellular antigens as well as to the immunobeads were carried out. Competition experiments showed that in the case of IgG MoAbs, the SAM beads bind predominantly to the Fc portion, whereas in the case of the IgM MoAbs, the Fab part plays a relatively greater role in the binding. The results indicate that if M-280 immunobeads are used, IgM MoAbs may profitably be included in antibody cocktails together with IgG antibodies in immunomagnetic purging of B-lymphoma cells. They suggest that in the case of cell bound MoAbs, the epitopes on IgG are more accessible to SAM beads than those of surface bound IgM molecules.(ABSTRACT TRUNCATED AT 250 WORDS)
Kvalheim
G
G
Institute for Cancer Research, Oslo, Norway.
Fjeld
J G
JG
Pihl
A
A
Funderud
S
S
Ugelstad
J
J
Fodstad
O
O
Nustad
K
K
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D001402:B-Lymphocytes / Q000276:immunology / Q000473:pathology*; D016026:Bone Marrow Transplantation; D002051:Burkitt Lymphoma / Q000276:immunology / Q000473:pathology*; D002469:Cell Separation / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D008280:Magnetics; D014407:Tumor Cells, Cultured / Q000276:immunology / Q000473:pathology
D000911:Antibodies, Monoclonal; D000912:Antibodies, Neoplasm; D007074:Immunoglobulin G; D007075:Immunoglobulin M