Direct measurement of antibody production in cell suspensions using an enzyme-linked immunosorbent assay.
We have developed an enzyme-linked immunosorbent assay (ELISA) to measure antibody production in cell suspensions using highly sensitive avidin-biotinylated peroxidase (ABC) reagents. The cells are serially diluted in media and placed directly on a standard antigen-coated microtiter plate for 6 h, and the plate is then washed and processed to determine the bound antibody. The amount of antibody detected is reduced by puromycin indicating in vitro synthesis. Standard errors of the means of optical density readings of replicate samples are less than 10%. The background readings and readings obtained using non-immune cells are negligible, demonstrating no significant contamination by endogenous peroxidases.
D000818:Animals; D000917:Antibody Formation / Q000187:drug effects; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007124:Immunoenzyme Techniques; D051379:Mice; D008813:Mice, Inbred ICR; D011691:Puromycin / Q000494:pharmacology
D011691:Puromycin