title: Structural mapping of immunoglobulin subclasses using multiplexed bead flow cytometry.
abstract: Monoclonal hybridomas secrete immunoglobulin molecules with a single specificity and distinct class/subclass structure. The determination of immunoglobulin structure can be used to facilitate hybridoma colony management and predict monoclonality. In this report, we used multiplexed bead flow cytometry to define hybridoma class/subclass. The assay was sufficiently sensitive to detect 50 ng/mL of antibody. The multiplexed bead assay efficiently defined traditional class/subclass determinants as well as more subtle patterns of crossreactivity. Further, the assay was combined with Poisson statistics to provide a numerical estimate of hybridoma monoclonality. The sensitivity and flexibility of this approach suggests the utility of multiplexed bead flow cytometry in the early management of immunoglobulin-secreting hybridomas.
authors.affiliation: Laboratory of Immunophysiology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Xiaoqun
authors.initials: X
authors.lastname: Jiang
authors.forename: Juan Pablo
authors.initials: JP
authors.lastname: Pratt
authors.forename: Harold O
authors.initials: HO
authors.lastname: Huss
authors.forename: Dino J
authors.initials: DJ
authors.lastname: Ravnic
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Mentzer
mesh_terms: D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D005434:Flow Cytometry; D006825:Hybridomas / Q000276:immunology; D007136:Immunoglobulins / Q000737:chemistry* / Q000145:classification*; D016130:Immunophenotyping; D051379:Mice; D008863:Microspheres; D010449:Peptide Mapping / Q000379:methods
chemical_list: D007136:Immunoglobulins