A rapid method for selecting specific hybridoma clones using paramagnetic Dynabeads.
This paper describes how specific hybridoma clones can be rapidly selected using paramagnetic beads coated with the antigen used for immunization. Spleen cells from a mouse immunized with fragment D dimer (DD) from plasminolysed fibrin were first fused with X-63 mouse myeloma cells. Paramagnetic monodisperse beads (precoated with sheep anti-mouse antibodies) were then coated with S4, a monoclonal antibody to DD, and subsequently with DD. Mixing such beads with the fused cells allowed selective harvesting of cells with membrane-expressed anti-DD gammaglobulins using a magnetic particle concentrator. Within 24 h, the cells spontaneously detached from the beads and were plated out on 96-well plates. Supernatants from the clones obtained were tested by the ELISA technique. Antibodies specific for DD were produced by 40-79% of the tested clones. It is concluded that it is possible to use antigen-coated paramagnetic beads to select, prior to cloning, hybridomas that produce specific antibodies. Implementation of this technique has significantly reduced costs and time in our efforts to obtain hybridoma clones of interest.
Bennick
A
A
Research Institute for Internal Medicine, Rikshospitalet, Oslo, Norway.
Brosstad
F
F
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D002999:Clone Cells; D004797:Enzyme-Linked Immunosorbent Assay; D005338:Fibrin Fibrinogen Degradation Products / Q000276:immunology; D006825:Hybridomas / Q000166:cytology*; D008280:Magnetics; D051379:Mice; D008807:Mice, Inbred BALB C
D005338:Fibrin Fibrinogen Degradation Products; C036309:fibrin fragment D