Secretion capture and report web: use of affinity derivatized agarose microdroplets for the selection of hybridoma cells.
A novel assay is described which allows the entrapment and detection of the immunoglobulin secreted from individual viable hybridoma cells using a secretion capture and reporter web (SCRW). By encapsulating the cells in agarose microdroplets which have been derivatized to create a fluorescent antigen-specific sandwich assay, flow cytometry can be used to identify and sort productive cells from a heterogeneous population. Using agarase, the cells can be recovered from the microdroplets and clonally expanded after selection. The assay has been used to reclone rare secretors from hybridoma cultures and to enhance the production of cultures with poor producers. The assay is easily generalized for the detection of any secreted protein for which specific antibodies or other ligands are available.
Gray
F
F
Syntex Discovery Research, Palo Alto, CA 94304, USA.
Kenney
J S
JS
Dunne
J F
JF
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D002214:Capsules; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006825:Hybridomas / Q000276:immunology*; D007158:Immunologic Techniques; D051379:Mice; D012685:Sepharose
D000911:Antibodies, Monoclonal; D002214:Capsules; D012685:Sepharose