title: Isolation of rare isotype switch variants in hybridoma cell lines using an agarose gel microdrop-based protein secretion assay.
abstract: Using gel microdrop (GMD) encapsulation technology and fluorescence-activated cell storing (FACS), we have developed a rapid, sensitive, and reliable method for discriminating and recovering rare isotypic switch variants in hybridoma cell lines. Using the GMD-based IgSwitch assay, a novel approach for isolating subpopulations of IgG-secreting hybridoma cells present at a frequency of approximately 1-10 in 10(6), we successfully isolated spontaneous and in vitro-induced isotypic switch variants in less than half the time required for conventional sublining. The effectiveness and specificity of the assay are demonstrated.
authors.affiliation: One Cell Systems, Inc, Cambridge, MA 02139, USA. yaks@onecell.com
authors.forename: Yevgenya
authors.initials: Y
authors.lastname: Akselband
authors.forename: Phillip T
authors.initials: PT
authors.lastname: Moen
authors.forename: Patricia
authors.initials: P
authors.lastname: McGrath
mesh_terms: D002469:Cell Separation / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D006825:Hybridomas / Q000145:classification / Q000166:cytology* / Q000378:metabolism*; D007074:Immunoglobulin G / Q000032:analysis* / Q000096:biosynthesis*; D011506:Proteins / Q000032:analysis / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D007074:Immunoglobulin G; D011506:Proteins