Transfer plate radioassay using cell monolayers to detect anti-cell surface antibodies synthesized by lymphocyte hybridomas.
A solid-phase [125I]Protein A radioassay for anti-cell surface antibodies is described which employs target cell monolayers cultured on fenestrated polyvinyl chloride 96-well plates ('transfer plates'). The calibrated aperture in the bottom of each well is small enough to retain fluid contents by surface tension during monolayer growth, but also permits fluid to enter the wells when transfer plates are lowered into receptacles containing washing buffer or test sera. To assay for antibodies directed against target cell surface antigens, transfer plates bearing monolayers are inserted into microculture plates with corresponding 96-well geometry, thereby simultaneously sampling 96 wells. This assay allows rapid screening of hundreds of hybird cell colonies for production of antibodies with desired tissue specificity.
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D002642:Chick Embryo; D006822:Hybrid Cells / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology; D051379:Mice; D009101:Multiple Myeloma; D011863:Radioimmunoassay / Q000379:methods*; D012160:Retina; D013205:Staphylococcal Protein A
D000906:Antibodies; D013205:Staphylococcal Protein A