Analysis of antibody reactivities in ELISA using protein blots as antigen substrates: S-ELISA.
We describe here a novel immunoassay procedure, designated strip-ELISA (S-ELISA), in which specific antigens are purified by SDS-PAGE, transferred to support membranes, and utilized in situ as substrate in routine ELISA procedures. Using two different lentivirus systems, simian immunodeficiency virus and equine infectious anemia virus, we demonstrate the utility of S-ELISA for screening hybridoma supernatants during production of monoclonal antibodies and for the dissection of polyclonal antibody responses into defined antigen specificities. The data presented here indicates that the S-ELISA combines the sensitivity of conventional ELISA with the specificity afforded by Western blotting procedures, producing a relatively simple and versatile assay for evaluating antibody reactivities.
Lo
W C
WC
Department of Biochemistry, Louisiana State University, Baton Rouge 70803.
Miller
M A
MA
Montelaro
R C
RC
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas; D007106:Immune Sera / Q000276:immunology; D004860:Infectious Anemia Virus, Equine / Q000276:immunology; D051379:Mice
D000914:Antibodies, Viral; D007106:Immune Sera