A simple in vitro method for raising monoclonal antibodies to cochlear proteins.
Monoclonal antibodies have been produced to mammalian hair cell antigens using a simple in vitro kit. Antigen was crudely prepared from dissected cochlear tissue by detergent extraction. There was no need to purify hair cells. Hybridoma supernatents were screened most efficiently on dissociated cells fixed with acetone. The immunisation method is sensitive to nanograms of antigen and can generate responses to conserved or weak antigens. The kit requires very little previous experience with cell culture and generates monoclonal antibodies within 3-4 weeks. It has overcome a number of problems with production of antibodies to hair cells and it should prove to be a very valuable tool in many laboratories.
Holley
M C
MC
Department of Physiology, Medical School, Bristol, UK.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000941:Antigens / Q000276:immunology; D015153:Blotting, Western; D002478:Cells, Cultured; D004586:Electrophoresis; D006168:Guinea Pigs; D006198:Hair Cells, Auditory / Q000276:immunology*; D007070:Immunoglobulin A / Q000276:immunology*; D051379:Mice; D008854:Microscopy, Electron; D013154:Spleen / Q000166:cytology / Q000276:immunology
D000911:Antibodies, Monoclonal; D000941:Antigens; D007070:Immunoglobulin A