A new and rapid method for the selection and cloning of antigen-specific hybridomas with magnetic microspheres.
A new and highly selective procedure is described for the rapid selection and cloning of antibody-secreting hybridomas with antigen-coated magnetic beads. Immune splenocytes were fused to Sp2 myeloma cells with a PEG/DMSO mixture. Cells were cultured in HAT and screened for the presence of specific antibody after 7-10 days. Hybridomas from the positive colonies were mixed with antigen-coated magnetic polymer beads and antigen-specific cells separated on a magnet. Cloning was carried out directly by limiting dilution with the magnetic beads still bound on the cells. No obvious toxic effects were observed. The antibodies established by this technique were of a high affinity (greater than 10(9) l/mol) and were generated in 50% of the usual process time. The procedure described here should greatly facilitate the process of obtaining hybridomas and expand the range of monoclonal antibodies available.
Horton
J K
JK
Amersham International plc, Forest Farm Industrial Estate, Cardiff, Wales, U.K.
Evans
O M
OM
Swann
K
K
Swinburne
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002459:Cell Fusion; D002469:Cell Separation / Q000379:methods*; D006063:Chorionic Gonadotropin / Q000276:immunology; D000939:Epitopes; D005260:Female; D006825:Hybridomas / Q000276:immunology; D007124:Immunoenzyme Techniques; D008280:Magnetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008863:Microspheres; D012397:Rosette Formation
D000911:Antibodies, Monoclonal; D006063:Chorionic Gonadotropin; D000939:Epitopes