A simple method for negative and positive selection of murine and human IgM-bearing lymphocytes based on the use of antibody-coated silica microparticles.
The use of octadecyl silica microparticles is proposed as an alternative method for negative and positive selection of IgM-bearing lymphocytes. SiC18 microparticles coated with affinity purified IgG anti-IgM were evaluated in their efficiency to deplete IgM-bearing cells. 6 mg of SiC18-IgG depleted an average of 97% IgM-bearing cells from a murine spleen cell suspension and of 84% IgM-bearing cells from human spleen cells. The viability of the selected cells was not affected by this procedure. We also analyzed the ability of whole serum to release IgM-bearing cells from the complex SiC18-target cell. The IgM releasing efficiency was 98%, with a resulting viability of 85%. These results indicate that SiC18 microparticles, saturated with specific antibodies, may be used as an alternative method for depletion or purification of cells bearing the corresponding antigen, from complex cell suspensions. The method is simple, efficient and inexpensive, representing an interesting alternative to the immunomagnetic selection.
Departamento de Medicina Preventiva Animal, Facultad de Ciencias Veterinarias y Pecuarias, Universidad de Chile, Santiago.
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Seeger
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Sepúlveda
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Vallejos
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JC
Aguillón
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Ferreira
D000818:Animals; D000906:Antibodies; D002469:Cell Separation / Q000379:methods*; D006801:Humans; D007075:Immunoglobulin M / Q000032:analysis*; D007158:Immunologic Techniques; D016131:Lymphocyte Subsets / Q000276:immunology*; D051379:Mice; D008863:Microspheres; D012680:Sensitivity and Specificity; D012822:Silicon Dioxide; D013154:Spleen / Q000166:cytology
D000906:Antibodies; D007075:Immunoglobulin M; D012822:Silicon Dioxide