A cell surface ELISA in the mouse using only poly-L-lysine as cell fixative.
An ELISA using plates coated with mouse spleen cells has been developed for analysis of antibodies to cell surface antigens. Such assays have been used extensively with human cells or with tumor cells in various species, but application to normal mouse lymphocytes has been limited. Use of normal spleen cells allows access to the genetic resources offered by recombinant, congenic, and mutant mouse strains, in the preparation of cell-coated ELISA plates, the use of glutaraldehyde was found to be unnecessary and it was eliminated, thereby avoiding the destruction of some cell surface determinants. Poly-L-lysine, which was used to treat plates, was found to provide sufficient adherence and preservation of the cells. Binding of biotinylated monoclonal antibodies to cells could be detected at approximately 10 ng/well. In inhibition assays, unlabeled antibodies could be detected at approximately 10 ng/well. Cell-coated plates are stable once prepared, and can be stored for months before use. The assay described can be used to quantitate levels of antibody to a particular epitope, and can also be adapted for screening of fusions for monoclonal antibodies to cell surface antigens.
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D000954:Antigens, Surface / Q000276:immunology*; D002448:Cell Adhesion; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006649:Histocompatibility Antigens / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D007518:Isoantibodies / Q000032:analysis; D051379:Mice; D008809:Mice, Inbred C3H; D010455:Peptides; D011107:Polylysine; D013154:Spleen / Q000276:immunology*
D000906:Antibodies; D000954:Antigens, Surface; D006649:Histocompatibility Antigens; D000949:Histocompatibility Antigens Class II; C024138:I-E-antigen; D007518:Isoantibodies; D010455:Peptides; D011107:Polylysine