ACSD labelling and magnetic cell separation: a rapid method of separating antibody secreting cells from non-secreting cells.
Several new ways of selecting cells have recently been developed. These include magnetic separation of cells by labelling with magnetic beads against the recombinant product, gel microdrop technology which encapsulates the cells in gelatine beads and matrix-based secretion assays. Affinity capture surface display (ACSD) is a matrix-based assay for the enrichment of high producing cells and relies on the strong affinity between biotin and avidin derivatives. Matrix-based assays have previously only been used for the enrichment of recombinant cells. Here, we have optimised this assay and developed a method of separating antibody producing cells from non-producing cells in a recombinant myeloma cell line using ACSD combined with MACS magnetic separation. The method is rapid, simple enough to become routine and adaptable to many different secreted products from recombinant mammalian cells.
Carroll
Silvia
S
Department of Chemical Engineering, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
Al-Rubeai
Mohamed
M
D000818:Animals; D000921:Antibody-Producing Cells / Q000737:chemistry / Q000276:immunology*; D001710:Biotin / Q000737:chemistry; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry; D046508:Culture Techniques; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D018189:Immunomagnetic Separation / Q000379:methods*; D051379:Mice; D008863:Microspheres; D019809:Streptavidin / Q000737:chemistry
D001710:Biotin; D019809:Streptavidin; D016650:Fluorescein-5-isothiocyanate