Isolation of rare isotype switch variants in hybridoma cell lines using an agarose gel microdrop-based protein secretion assay.
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.
Akselband
Yevgenya
Y
One Cell Systems, Inc, Cambridge, MA 02139, USA. yaks@onecell.com
Moen
Phillip T
PT
McGrath
Patricia
P
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
D007074:Immunoglobulin G; D011506:Proteins